人羊膜上皮细胞横向分化为肝细胞样细胞及脾内移植的初步研究
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摘要
在胚胎发育过程中,肝脏的整个发育过程是一个涉及多基因、多环节、多途径的网络调控过程。1965年Farber的研究揭开了对于肝细胞研究的新篇章,他首次提出在肝内存在小上皮细胞,即卵圆细胞。他的发现推动了肝再生及肝细胞肝癌发病机制的深入研究。目前认为肝干细胞(hepaticstem cell,HSC)来源于前肠内胚层,在胚胎发育过程中以未成熟的肝细胞的形式存在,其形态与胆管上皮细胞相似,其生化特征类似于胚胎干细胞。20世纪末,大量研究表明,成体干细胞可以跨越胚层限制分化为肝细胞。美国科学家近年宣布,他们在羊水中发现了和胚胎干细胞一样有效的干细胞,并在实验室中使这种干细胞分化成肌肉、骨头、脂肪、血管、神经和肝脏细胞。和胚胎干细胞相比,这种干细胞更容易分化成各种组织,且不会形成一种名为畸胎瘤的良性肿瘤。来源于人废弃胎盘的羊膜上皮细胞(human amnioticepithelial cells,hAECs)是这样一类极具研究潜力的细胞。目前国内外这方面的研究尚处于起步阶段。
本课题对人羊膜上皮细胞完成了体外检测和鉴定,对其向肝细胞样细胞体外分化的条件进行了摸索和优化,在成功诱导生成肝细胞样细胞的基础上,将诱导获得的肝细胞样细胞移植入裸小鼠体内,检测了诱导获得的肝细胞样细胞是否具有生物学功能。本研究分为3个部
目的:建立人羊膜上皮细胞的体外分离方法及培养体系,检测人羊膜上皮细胞的生物学特性,检测人羊膜上皮细胞的AAV-EGFP感染效率。
方法:使用胰蛋白酶从人羊膜上消化人羊膜上皮细胞进行培养,细胞免疫荧光检测胚胎干细胞表面SSEA-1、SSEA-3、SSEA-4和TRA-1-60、TRA-1-81标记;RT-PCR检测全能性相关基因TERF、THY1、LEFTYA、Rex-1、Sox-2、Oct-4和nanog等的表达;腺相关病毒感染检测人羊膜上皮细胞的AAV-EGFP感染效率。
结果:人羊膜上皮细胞表达SSEA3、SSEA4、TRA-1-60、TRA-1-81等胚胎干细胞的特异性标记;表达TERF1、THY、EFTYA、Rex-1、SOX2、Oct-4和Nanog等全能性相关基因;人羊膜上皮细胞的AAV-GFP感染效率在病毒浓度为10~(-3)pfu/ml时可达58.2%±1.3%。
结论:建立了hAECs的细胞分离方法及适合hAECs生长和增殖的体外培养体系;hAECs具干细胞特征;体外培养的hAECs细胞可以被AAV-GFP感染,并表达GFP,且表达效率达50%以上。
目的:研究Dex,HGF,IGF等细胞因子联合使用的诱导体系对hAECs向肝细胞样(hepatocyte-like)细胞诱导分化的影响。
方法:采用Dex,HGF,IGF等细胞因子联合使用方法诱导培养hAECs向肝细胞样细胞分化,诱导周期为两周。诱导过程中采用RT-PCR鉴定细胞ALB、CYP1A1、CYP1A2、IGFR、c-met等肝细胞相关关键功能基因的表达和HNF3、HNF4和C/EBPa三种转录因子的表达。流式细胞术分析集落细胞表面标记ALB、AFP和CK18的时程变化;为了优化诱导体系,检测了Dex、HGF、IGF等细胞因子的剂量依赖性;比较不同代数的hAECs在诱导向肝细胞样细胞分化的ALB基因表达情况。
结果:在诱导两周后的hAECs中可以检测到ALB、CYP1A1、CYP1A2、IGFR、c-met等肝细胞相关关键功能基因的表达,且这些功能基因的表达呈现逐渐递增的趋势。提示hAECs可能已经被成功诱导为肝细胞样细胞。诱导前的hAECs表达HNF1(一种肝细胞转录因子),不表达HNF3、HNF4和C/EBPa;而诱导后的hAECs不仅可以表达HNF1,还可以表达HNF3,HNF4和C/EBPa,并且HNF1的表达在诱导前后的hAECs中也有所不同,诱导后的hAECs中HNF1的表达水平明显高于没有经过诱导的hAECs。细胞表面标记检测发现:诱导6天时,hAECs主要表达AFP+,约为15.1±2.1%;随着诱导时间的延长,10天时hAECs表达AFP+/ALB+,约为6.5±1.4%;而诱导14天时,hAECs基本上只表达ALB+,为13.9±2.3%。诱导10天时的hAECs开始表达ALB+/CKl8+;约为2.5±1.4%;诱导14天时,细胞依然表达ALB+/CK18+,且这种双阳性的细胞量有明显增加,为18.9±3.1%;同时CK18+细胞数并没有明显减少,诱导10天时,CK18+细胞数为16.1±1.2%;诱导14天时,CK18+细胞数为21.3±4.6%。提示,诱导过程中的hAECs随着诱导时间的延长有一个逐渐成熟的过程。剂量依赖性实验结果显示,IGF、HGF均存在剂量依赖性,而Dex的剂量依赖性实验结果不具有统计学意义,认为不存在剂量依赖性。对不同代数的hAECs在诱导向肝细胞样细胞分化的ALB基因表达情况进行比较,结果显示,传代三代的hAECs诱导后均能表达ALB基因,且在表达强度上没有明显区别
结论:hAECs确实可以在体外向肝细胞样细胞进行诱导分化,并能被成功诱导分化为有功能的肝细胞样细胞;hAECs向肝细胞样细胞的诱导过程中,对于HGF和IGF具有剂量依赖性,而对于Dex的这种剂量依赖性并不明显;传代三代内的hAECs诱导后,在肝细胞样细胞特异性功能基因ALB的表达上没有明显区别。
目的:观察人羊膜上皮细胞(hAECs)在肝脏受损裸鼠脾内移植后是否具有肝细胞样功能的表达并探讨其治疗肝功能损伤的可行性。
方法: 40只裸小鼠,随机分为三组,A组:20只,行2/3肝叶切除后,自脾下极移植5×10~6绿色荧光蛋白标记的hAECs;B组:10只,不行肝叶切除,自脾下极移植5×10~6绿色荧光蛋白标记的hAECs;C组:10只,行2/3肝叶切除后,自脾下极注射生理盐水0.2 mL。荧光显微镜hAECs体内示踪并观察其是否表达肝细胞特异性蛋白;术后2周A、B组各10只裸小鼠采血定量检测裸鼠血中人白蛋白及肝功能,术后4周A组另10只及C组10只裸小鼠做以上相同检测。
结果: A组镜下肝脏和脾脏组织中均发现绿色荧光信号的hAECs,大部分表达人白蛋白;B、C组镜下均未见绿色荧光细胞。A组血中可检测到人白蛋白且术后4周明显高于术后2(3.9±1.34vs0.37±0.14,P<0.01);术后4周A组肝功能(ALB、ALT、AST)明显优于C组(p<0.05),A组肝功能术后4周亦优于术后2周(p＜0.05)。
结论:肝损伤可以有效诱导hAECs向肝迁移增殖,大部分hAECs在肝细胞损伤的微环境下有肝细胞样功能表达,hAECs移植可以在一定程度上修复受损肝功能。
In the process of embryonic development, the development of the liver is involved in a multi-gene, multi-link, multi-channel network and the hepatic stem cells play a very important role in this process. This kind of stem cells was developed from the foregut. it is similar to bile duct epithelial cell in morphology and to embryonic stem cells in biochemical.
In the end of the 20th century, numerous studies show that trandifferentiation could happen when adult stem cells were cultured in special condition. In recent years, American scientists announced that they found some stem cells in the amniotic fluid as effective as embryonic stem cells, and they found that those stem cells can differentiated into muscle, bone, fat, blood vessels, nerves and liver cells in the laboratory. Compared to embryonic stem cells, they could differentiate into various organizations more easily, and could not formate teratomas. Human amniotic epithelial cells, extracted from the abandoned placenta is such a great research potentiality cells. Current domestic and international research in this area is still in its infancy.
The aim of theasis was to discuss the method of isolation and identification of the human amniotic epithelial cells, to invstigate the posibility of its transdifferentiation into hepatocyte-like cells and the Intrasplenic Transplantation. This study is divided into three parts, the main research methods and results are as follows:
Objectives: To isolate hAECs from amnion and establish a culture system for hAECs in vitro. To identify the stem cell related characteristics of these isolated hAECs . To test the AAV transfection efficiency of hAECs.
Methods: Trypsin-EDTA was used to digest the amnions we collected. The cell sample of P1 passage was collected and then semi-quantity RT-PCR was performed to examine the pluripotency-related genes expression, such as TERF、THY1、LEFTYA、Rex-1、Sox-2、Oct-4 and nanog. Immunocytofluorescental analysis was taken to test the expression of stem cell related markers, such as SSEA-K SSEA-3、SSEA-4 and TRA-1-60、TRA-1-81. The AAV transfection efficiency to hAECs was analyzed with AAV-GFP in vitro.
Results: RT-PCR showed that TERF、THY1、LEFTYA、Rex-1、Sox-2、Oct-4 and nanog were expressed in P1 hAECs. Immunocytofluorescent analysis showed that most of hAECs were SSEA-3~+,SSEA-4~+,TRA-1-60~+ and TRA-1-81~+. The transfection efficiency of hAECs estimated by EGFP staining was about 58.2%±1.3 % with AAV concentration of 10~(-3) pfu/ml.
Conclusions: We isolated hAECs from amnions successfully and stablished a culture system for hAECs in vitro. hAECs was a kind of stem cells. hAECs cultured in vitro can express GFP and its expression efficiency is more than 50 percent.
Objective: To establish a optimizing culture system of the inducing process and study the contribution of the three factors in the system: Dex, HGF, IGF. To discuss detecting index of the hepatic transdiffernetion of HAECs.
Methods: The induction cycle lasted for two weeks, in the end of induction functional genes and transcriptional factors were selected to detect the hepatic fate commitment. For example, the expression of albumin, CYP1A1, CYP1A2, IGFR, c-met, and HNF3, HNF4 and C/EBP were examined by the RT-PCR. Then flow cytometric analysis were performed to test the cell surface markers such as ALB, AFP and CK18 to explore the hepatocyte-like cell maturation; In order to optimize the induction system, detected the dose-dependent of the Dex, HGF, IGF cytokines; Albumin gene expression of different generation's amniotic epithelial cells were examed in the processes of the amniotic epithelial cells differentiated into liver-like cell.
Results: Two weeks after the induction, the expression of albumin, CYP1A1, CYP1A2, IGFR, c-met and other liver cell-related key functional gene's can be detected in the human amniotic epithelial cells. Further study found that these gene's expression level showed the trend of gradual increasement which suggested that hAECs may have successfully be induced into hepatocyte-like cells. After inducement, HAECs expressed HNF3, HNF4 and C/ EBPa which were not detected before the induction; and the expression level of HNF1 is significantly higher than that not been induced. The detection of cell surface markers found that: 6 days after be induced, hAECs mainly were stained AFP+, the positive rate was about 15.1±2.1% ; with the time extension, 10 days after be induced, part of the hAECs showed AFP+/ALB+, which was about 6.5±1.4%; and on 14 days, hAECs basically only showed ALB+, the rate was about 13.9±2.3%; the positive rate of the ALB+ cells increasing indicated a grandual founctional maturation from the hAECs to hepatocyte-like cells. Similaritly, the CK18+ cells in the whole population were also increased in the number, for example on the 10 day, the rate was 16.1±1.2%; on the 14~(th) day, 21.3±4.6%, which proved the above hypothesis of the trandifferentiation. Experimental results of the dose-dependent study show that: IGF, HGF were dose dependent,while Dex's dose-dependent experimental results was not statistically significant, suggested that dose-dependent manner does not exist. Three passages of HAECs were compared in the gene expression of albumin during the induction. It showed that all the third generation cells which were induced, could all expressed the albumin gene and the expression had no obvious distinction.
Conclusion: hAECs could indeed been induced to differentiate into hepatocyte-like cells in vitro, and could be successfully induced to form special functional hepatocyte-like cells; In the induction process, had a dose-dependent on HGF and IGF, whereas this dose-dependent is not obvious on Dex; In the passage of three generations, the expression of albumin,which was the hepatocyte -like-cells-specific gene, had not significantly difference.
Objective To observe whether human Amniotic Epithelial Cells (hAECs) have the potential to possess hepatocyte-like function after intrasplenic transplantation in nude mice with liver injury as well as the therapeutic feasibility to repair damaged liver.
Methods 40 nude mice were randomly divided into three groups: Group A ( n=20) : 5×10~6hAECs labeled with green fluorescent protein were transplanted into the spleen of nude mice undergone 2/3 hepatectomized; Group B: treated with same cells as in A,but without hepatectomized; Group C: 0.2 saline was injected into the spleen of nude mice undergone 2/3 hepatectomized. The hAECs were detected with immunofluorescent microscope and their albumin expression were studied. Two and four weeks after the cell transplantation, Liver function and quantitative human albumin were measured respectively in all groups.
Results The hAECs labeled green fluorescent protein were observed in liver and spleen tissue in group A and mostly expressed albumin.While in group B,C ,there is no such findings. The human albumin was detectable in serum of group A .The level of serum human albumin in group A in 4 weeks after transplantation was higher than that in 2 weeks after transplantation (3.9±1.34vs0.37±0.14, P<0.01). Four weeks after transplantation, the liver functions in Group A were restored significantly compared with those of Group C,The liver functions in Group A in 4 weeks after transplantation were improved significantly compared with those in 2 weeks after transplantation while the level of ALT was also different between two groups(121.2±20.8 vs. 191.8±28.9, P＜0.05).
Conclusion The stimulus of liver damage might enhances the migration of hAECs to the liver ,in which a majority of the hAECs have the potential to possess hepatocyte-like function in nude mice . Transplantation of hAECs might amend the damaged liver function to a certain extent.
本课题对人羊膜上皮细胞完成了体外检测和鉴定,对其向肝细胞样细胞体外分化的条件进行了摸索和优化,在成功诱导生成肝细胞样细胞的基础上,将诱导获得的肝细胞样细胞移植入裸小鼠体内,检测了诱导获得的肝细胞样细胞是否具有生物学功能。本研究分为3个部
目的:建立人羊膜上皮细胞的体外分离方法及培养体系,检测人羊膜上皮细胞的生物学特性,检测人羊膜上皮细胞的AAV-EGFP感染效率。
方法:使用胰蛋白酶从人羊膜上消化人羊膜上皮细胞进行培养,细胞免疫荧光检测胚胎干细胞表面SSEA-1、SSEA-3、SSEA-4和TRA-1-60、TRA-1-81标记;RT-PCR检测全能性相关基因TERF、THY1、LEFTYA、Rex-1、Sox-2、Oct-4和nanog等的表达;腺相关病毒感染检测人羊膜上皮细胞的AAV-EGFP感染效率。
结果:人羊膜上皮细胞表达SSEA3、SSEA4、TRA-1-60、TRA-1-81等胚胎干细胞的特异性标记;表达TERF1、THY、EFTYA、Rex-1、SOX2、Oct-4和Nanog等全能性相关基因;人羊膜上皮细胞的AAV-GFP感染效率在病毒浓度为10~(-3)pfu/ml时可达58.2%±1.3%。
结论:建立了hAECs的细胞分离方法及适合hAECs生长和增殖的体外培养体系;hAECs具干细胞特征;体外培养的hAECs细胞可以被AAV-GFP感染,并表达GFP,且表达效率达50%以上。
目的:研究Dex,HGF,IGF等细胞因子联合使用的诱导体系对hAECs向肝细胞样(hepatocyte-like)细胞诱导分化的影响。
方法:采用Dex,HGF,IGF等细胞因子联合使用方法诱导培养hAECs向肝细胞样细胞分化,诱导周期为两周。诱导过程中采用RT-PCR鉴定细胞ALB、CYP1A1、CYP1A2、IGFR、c-met等肝细胞相关关键功能基因的表达和HNF3、HNF4和C/EBPa三种转录因子的表达。流式细胞术分析集落细胞表面标记ALB、AFP和CK18的时程变化;为了优化诱导体系,检测了Dex、HGF、IGF等细胞因子的剂量依赖性;比较不同代数的hAECs在诱导向肝细胞样细胞分化的ALB基因表达情况。
结果:在诱导两周后的hAECs中可以检测到ALB、CYP1A1、CYP1A2、IGFR、c-met等肝细胞相关关键功能基因的表达,且这些功能基因的表达呈现逐渐递增的趋势。提示hAECs可能已经被成功诱导为肝细胞样细胞。诱导前的hAECs表达HNF1(一种肝细胞转录因子),不表达HNF3、HNF4和C/EBPa;而诱导后的hAECs不仅可以表达HNF1,还可以表达HNF3,HNF4和C/EBPa,并且HNF1的表达在诱导前后的hAECs中也有所不同,诱导后的hAECs中HNF1的表达水平明显高于没有经过诱导的hAECs。细胞表面标记检测发现:诱导6天时,hAECs主要表达AFP+,约为15.1±2.1%;随着诱导时间的延长,10天时hAECs表达AFP+/ALB+,约为6.5±1.4%;而诱导14天时,hAECs基本上只表达ALB+,为13.9±2.3%。诱导10天时的hAECs开始表达ALB+/CKl8+;约为2.5±1.4%;诱导14天时,细胞依然表达ALB+/CK18+,且这种双阳性的细胞量有明显增加,为18.9±3.1%;同时CK18+细胞数并没有明显减少,诱导10天时,CK18+细胞数为16.1±1.2%;诱导14天时,CK18+细胞数为21.3±4.6%。提示,诱导过程中的hAECs随着诱导时间的延长有一个逐渐成熟的过程。剂量依赖性实验结果显示,IGF、HGF均存在剂量依赖性,而Dex的剂量依赖性实验结果不具有统计学意义,认为不存在剂量依赖性。对不同代数的hAECs在诱导向肝细胞样细胞分化的ALB基因表达情况进行比较,结果显示,传代三代的hAECs诱导后均能表达ALB基因,且在表达强度上没有明显区别
结论:hAECs确实可以在体外向肝细胞样细胞进行诱导分化,并能被成功诱导分化为有功能的肝细胞样细胞;hAECs向肝细胞样细胞的诱导过程中,对于HGF和IGF具有剂量依赖性,而对于Dex的这种剂量依赖性并不明显;传代三代内的hAECs诱导后,在肝细胞样细胞特异性功能基因ALB的表达上没有明显区别。
目的:观察人羊膜上皮细胞(hAECs)在肝脏受损裸鼠脾内移植后是否具有肝细胞样功能的表达并探讨其治疗肝功能损伤的可行性。
方法: 40只裸小鼠,随机分为三组,A组:20只,行2/3肝叶切除后,自脾下极移植5×10~6绿色荧光蛋白标记的hAECs;B组:10只,不行肝叶切除,自脾下极移植5×10~6绿色荧光蛋白标记的hAECs;C组:10只,行2/3肝叶切除后,自脾下极注射生理盐水0.2 mL。荧光显微镜hAECs体内示踪并观察其是否表达肝细胞特异性蛋白;术后2周A、B组各10只裸小鼠采血定量检测裸鼠血中人白蛋白及肝功能,术后4周A组另10只及C组10只裸小鼠做以上相同检测。
结果: A组镜下肝脏和脾脏组织中均发现绿色荧光信号的hAECs,大部分表达人白蛋白;B、C组镜下均未见绿色荧光细胞。A组血中可检测到人白蛋白且术后4周明显高于术后2(3.9±1.34vs0.37±0.14,P<0.01);术后4周A组肝功能(ALB、ALT、AST)明显优于C组(p<0.05),A组肝功能术后4周亦优于术后2周(p＜0.05)。
结论:肝损伤可以有效诱导hAECs向肝迁移增殖,大部分hAECs在肝细胞损伤的微环境下有肝细胞样功能表达,hAECs移植可以在一定程度上修复受损肝功能。
In the process of embryonic development, the development of the liver is involved in a multi-gene, multi-link, multi-channel network and the hepatic stem cells play a very important role in this process. This kind of stem cells was developed from the foregut. it is similar to bile duct epithelial cell in morphology and to embryonic stem cells in biochemical.
In the end of the 20th century, numerous studies show that trandifferentiation could happen when adult stem cells were cultured in special condition. In recent years, American scientists announced that they found some stem cells in the amniotic fluid as effective as embryonic stem cells, and they found that those stem cells can differentiated into muscle, bone, fat, blood vessels, nerves and liver cells in the laboratory. Compared to embryonic stem cells, they could differentiate into various organizations more easily, and could not formate teratomas. Human amniotic epithelial cells, extracted from the abandoned placenta is such a great research potentiality cells. Current domestic and international research in this area is still in its infancy.
The aim of theasis was to discuss the method of isolation and identification of the human amniotic epithelial cells, to invstigate the posibility of its transdifferentiation into hepatocyte-like cells and the Intrasplenic Transplantation. This study is divided into three parts, the main research methods and results are as follows:
Objectives: To isolate hAECs from amnion and establish a culture system for hAECs in vitro. To identify the stem cell related characteristics of these isolated hAECs . To test the AAV transfection efficiency of hAECs.
Methods: Trypsin-EDTA was used to digest the amnions we collected. The cell sample of P1 passage was collected and then semi-quantity RT-PCR was performed to examine the pluripotency-related genes expression, such as TERF、THY1、LEFTYA、Rex-1、Sox-2、Oct-4 and nanog. Immunocytofluorescental analysis was taken to test the expression of stem cell related markers, such as SSEA-K SSEA-3、SSEA-4 and TRA-1-60、TRA-1-81. The AAV transfection efficiency to hAECs was analyzed with AAV-GFP in vitro.
Results: RT-PCR showed that TERF、THY1、LEFTYA、Rex-1、Sox-2、Oct-4 and nanog were expressed in P1 hAECs. Immunocytofluorescent analysis showed that most of hAECs were SSEA-3~+,SSEA-4~+,TRA-1-60~+ and TRA-1-81~+. The transfection efficiency of hAECs estimated by EGFP staining was about 58.2%±1.3 % with AAV concentration of 10~(-3) pfu/ml.
Conclusions: We isolated hAECs from amnions successfully and stablished a culture system for hAECs in vitro. hAECs was a kind of stem cells. hAECs cultured in vitro can express GFP and its expression efficiency is more than 50 percent.
Objective: To establish a optimizing culture system of the inducing process and study the contribution of the three factors in the system: Dex, HGF, IGF. To discuss detecting index of the hepatic transdiffernetion of HAECs.
Methods: The induction cycle lasted for two weeks, in the end of induction functional genes and transcriptional factors were selected to detect the hepatic fate commitment. For example, the expression of albumin, CYP1A1, CYP1A2, IGFR, c-met, and HNF3, HNF4 and C/EBP were examined by the RT-PCR. Then flow cytometric analysis were performed to test the cell surface markers such as ALB, AFP and CK18 to explore the hepatocyte-like cell maturation; In order to optimize the induction system, detected the dose-dependent of the Dex, HGF, IGF cytokines; Albumin gene expression of different generation's amniotic epithelial cells were examed in the processes of the amniotic epithelial cells differentiated into liver-like cell.
Results: Two weeks after the induction, the expression of albumin, CYP1A1, CYP1A2, IGFR, c-met and other liver cell-related key functional gene's can be detected in the human amniotic epithelial cells. Further study found that these gene's expression level showed the trend of gradual increasement which suggested that hAECs may have successfully be induced into hepatocyte-like cells. After inducement, HAECs expressed HNF3, HNF4 and C/ EBPa which were not detected before the induction; and the expression level of HNF1 is significantly higher than that not been induced. The detection of cell surface markers found that: 6 days after be induced, hAECs mainly were stained AFP+, the positive rate was about 15.1±2.1% ; with the time extension, 10 days after be induced, part of the hAECs showed AFP+/ALB+, which was about 6.5±1.4%; and on 14 days, hAECs basically only showed ALB+, the rate was about 13.9±2.3%; the positive rate of the ALB+ cells increasing indicated a grandual founctional maturation from the hAECs to hepatocyte-like cells. Similaritly, the CK18+ cells in the whole population were also increased in the number, for example on the 10 day, the rate was 16.1±1.2%; on the 14~(th) day, 21.3±4.6%, which proved the above hypothesis of the trandifferentiation. Experimental results of the dose-dependent study show that: IGF, HGF were dose dependent,while Dex's dose-dependent experimental results was not statistically significant, suggested that dose-dependent manner does not exist. Three passages of HAECs were compared in the gene expression of albumin during the induction. It showed that all the third generation cells which were induced, could all expressed the albumin gene and the expression had no obvious distinction.
Conclusion: hAECs could indeed been induced to differentiate into hepatocyte-like cells in vitro, and could be successfully induced to form special functional hepatocyte-like cells; In the induction process, had a dose-dependent on HGF and IGF, whereas this dose-dependent is not obvious on Dex; In the passage of three generations, the expression of albumin,which was the hepatocyte -like-cells-specific gene, had not significantly difference.
Objective To observe whether human Amniotic Epithelial Cells (hAECs) have the potential to possess hepatocyte-like function after intrasplenic transplantation in nude mice with liver injury as well as the therapeutic feasibility to repair damaged liver.
Methods 40 nude mice were randomly divided into three groups: Group A ( n=20) : 5×10~6hAECs labeled with green fluorescent protein were transplanted into the spleen of nude mice undergone 2/3 hepatectomized; Group B: treated with same cells as in A,but without hepatectomized; Group C: 0.2 saline was injected into the spleen of nude mice undergone 2/3 hepatectomized. The hAECs were detected with immunofluorescent microscope and their albumin expression were studied. Two and four weeks after the cell transplantation, Liver function and quantitative human albumin were measured respectively in all groups.
Results The hAECs labeled green fluorescent protein were observed in liver and spleen tissue in group A and mostly expressed albumin.While in group B,C ,there is no such findings. The human albumin was detectable in serum of group A .The level of serum human albumin in group A in 4 weeks after transplantation was higher than that in 2 weeks after transplantation (3.9±1.34vs0.37±0.14, P<0.01). Four weeks after transplantation, the liver functions in Group A were restored significantly compared with those of Group C,The liver functions in Group A in 4 weeks after transplantation were improved significantly compared with those in 2 weeks after transplantation while the level of ALT was also different between two groups(121.2±20.8 vs. 191.8±28.9, P＜0.05).
Conclusion The stimulus of liver damage might enhances the migration of hAECs to the liver ,in which a majority of the hAECs have the potential to possess hepatocyte-like function in nude mice . Transplantation of hAECs might amend the damaged liver function to a certain extent.
引文
[1] Strain AJ. C rosby H A. Hepatic stem cells.Gut,2000.46:743-745.
[2] Sanchez-Ramos JR. Neural cells derived from adult bone marrow and umbilicalco rdb lood.J N eurosciR es.2002,6 9(6):880-93.
[3] Theise ND, Badve S, Saxena et al. Derivation of hepatocytes from bonem arrowc ellsin m icea fterra diation-inducedm yeloablation.H epatology, 2000,3 l(1):235-240.
[4] Theise N D, N immakayalu M , Gardner R, et al . Liver from bone marrow In humans. Hepatology,2000,32:11-14.
[5] Thalheimcr U. Capre F..Liver transplantation: making the best out of what we Have Dig Dis Sci,2002,47(5):945-953.
[6] Stockrnanl HB, Ijxermans JN.Prospects for the treatment of acute liver Failure. Eur J Gastroenterol Hepatol,2002,14(2):195-203.
[7] Fox IJ,Chowdhury JR,Kaufman SS,et al. Treatment of the Crigler-Najjar syndrome type 1 with hepatocyte transplantation.N Engl J Med. 1998;338(20): 1422-1426.
[8] Lorenti AS. Hepatic stem cells. Medicina (B Aires).2001;61(sptl):614-620.
[9] Kobayashi N Fuliwara T, Wcsterman KA. et al. Prevention of acute liver failure in rats with reversibly immortalized human hepatocytes. Science. 2000.287(19): 1258-1262.
[10] Cogle CR ,Yachnis AT,Laywell ED, et al. Bone marrow transdifferentiation in brain after transplantation: a retrospective study. Lancet, 2004, 363 : 1432-1437.
[11] Schwartz RE, Reyes M, Koodie L, et al. Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. J Clin Invest, 2002, 109: 1291-1302.
[12] Spees JL, Olson SD, Ylostalo J, et al. Differentiation, cell fusion, and nuclear fusion during ex vivo repair of epithelium by human adult stem cells from bone marrow stroma. Proc Natl Acad Sci USA, 2003, 100: 2397.
[13] 王 萍。 中华肝脏病杂志, 2004, 1 2 : 718
[14] SakuragawaN ,T ohyamaJ ,Y amamotoH . Immunostaining of human amniotic epithelil cells:possible use as a transgenec arrier in gene therapy for in bonrerrors of metabolism.Cell Transplant. 1995;4(3):343-346.
[15] Hunt JS, W ood GW. Interferon-gamma induces class I HLA and beta 2-microg lobulin expression by human amnion cells.J I mmunol.l 986;136(2):364- 367.
[16] Tabei I, Hashimoto H , Ishiwata I, et al . Characteristics of hepatocytes derived from early ES cells and treatment of surgically induced liver failure rats by transplantation. Transplant Proc , 2005 , 37 (1) :262-264.
[17] Nishitai R , Koch CA , Ogata K, et al . Toward the survival and function of xenogeneic hepatocyte grafts. Liver Transpl ,2005 , 11 (1) :39 - 50.
[18] Gunsalus JR,Brady DA,Coulter SM, et al . Reduction of serum cholesterol in Watanabe rabbits by xenogeneic hepatocellular transplantation. Nat Med , 1997 , 3(1) :48-53.
[19] Nagata H , Ito M, Cai J , et al. Treatment of cirrhosis and liver failure in rats by hepatocyte xenotransplantation. Gastroenterology, 2003 , 124(2) :422-431.
[20] Maruyama M, Totsugawa T , Kunieda T , et al . Hepatocyte isolation and transplantation in the pig. Cell Transplant, 2003 , 12(6) :593 -598.
[21] Bilir BM, Guinette D , Karrer F , et al . Hepatocyte transplantation in acute liver failure. Liver Transpl, 2000 , 6 (1) :32-40.
[22] Fisher RA , Bu D , Thompson M, et al . Defining hepatocellular chimerism in a liver failure patient bridged with hepatocyte infusion. Transplantation , 2000 , 69 (2):303-307.
[23] Strom SC , Fisher RA , Thompson M, et al . Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure. Transplantation , 1997,63 (4) :559-569.
[24] Nakamura J , Okamoto T , Schumacher IK, et al . Treatment of surgically induced acute liver failure by transplantation of conditionally immortalized hepatocytes. Transplantation , 1997 , 63(11):1541 - 1547.
[25] Cai J , Ito M, Nagata H , et al. Treatment of liver failure in rats with end-stage cirrhosis by transplantation of immortalized hepatocytes. Hepatology , 2002 , 36(2) :386 -394.
[26] Koyano S, Fukui A, Uchida S, Yamada K, et al. Synthesis and release of activin and noggin by cultured human amniotic epithelial cells. Dev Growth Differ. 2002; 44(2): 103-112.
[27] Sakuragawa N, Misawa H, Ohsugi K, et al. Evidence for active acetylcholine metabolism in human amniotic epithelial cells: applicable to intracerebral allografting for neurologic disease. Neurosci Lett. 1997; 232(l):53-56.
[28] Sankar V, Muthusamy R. Role of human amniotic epithelial cell transplantation in spinal cord injury repair research. Neuroscience. 2003; 118(1):11-17.
[29] Tseng SC, Meller D, Anderson DF, et al. Ex vivo preservation and expansion of human limbal epithelial stem cells on amniotic membrane for treating corneal diseases with total limbal stem cell deficiency. Adv Exp Med Biol. 2002; 506:1323-1334.
[30] akuragawa N, Tohyama J, Yamamoto H. Immunostaining of human amniotic epithelial cells: possible use as a transgene carrier in gene therapy for inborn errors of metabolism. Cell Transplant. 1995; 4(3):343-346.
[31] Sakuragawa N, Enosawa S, Ishii T, et al. Human amniotic epithelial cells are promising transgene carriers for allogeneic cell transplantation into liver. J Hum Genet. 2000; 45 (3): 171-176.
[32] Bhattacharya B, Miura T, Brandenberger R, et al. Gene expression in human embryonic stem cell lines: unique molecular signature. Blood. 2004; 103 (8): 2956-64.
[33] Chambers I, Colby D, Robertson M, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 2003; 113 (5): 643-655.
[34] Cavaleri F, Scholer HR. Nanog: a new recruit to the embryonic stem cell orchestra. Cell. 2003; 113 (5): 551-552.
[35] Brivanlou AH, Gage FH, Jaenisch R, et al.Stem cells. Setting standards for human embryonic stem cells.Science. 2003; 300(5621):865.
[36] Gelse K, von der Mark K. Cartilage regeneration by gene therapy.Curr Gene Ther,2003; 3: 305-317.
[37] Coffin RS, Thomas SK, Thomas NSB. Pure populations of transduced Primary human cells can be produced using GFP expression herpes virus vectors and flow cytometry. Gene Ther, 1998; 5: 718-722.
[38] Soderqvist H , Imerch G, Kihlark M. Intracellular distribution of an integral nuclear pore membrane protein fused to green fluorescent protein locatization of a targeting domain. Eur J Biochem,1997; 250(3) :808-813.
[39] Kalajzic I, Braut A, Guo D, Jiang X, Kronenberg MS, Mina M, Harris MA, Harris SE, Rowe DW, Dentin matrix protein 1 expression during osteoblastic differentiation, generation of an osteocyte GFP transgene . Bone, 2004; 35(1):74-82.28
[40] Toshio Miki,a Thomas Lehmann,a Hongbo Cai,a Donna B. Stolz,b,c Stephen C. Stroma,cStem Cell Characteristics of Amniotic Epithelial Cells.Stem Cells 2005;23:1549 - 1559.
[41 ] Ilancheran S; Michalska A; Peh G; Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9, EPUB .
[42] 裴雪涛,刘大庆.干细胞. 组织器官重建的理想种子细胞. 中国修复重建外科杂志, 2006, 20 (4): 344-348.
[43] Strick2M archand H, Mo ro san S, Charneau P, et al. Bipo tential mouse embryonic liver stem cell lines contribute to liver regeneration and differentiate as bile ducts and hepatocytes. P roc N atl A cad SciU SA, 2004, 101 (22): 8360-8365.
[44] Zheng YW , Taniguch i H. D iversity of hepatic stem cells in the fetal and adult liver. Sem in L iver D is, 2003, 23 (4): 3372-3480.
[45] Paku S, Schnur J , N agy P, et al. O rigin and structural evo lution of the early p ro liferating oval cells in rat liver. Am J Patho 1, 2001, 158 (4): 1313-1323.
[46] He ZP, TanWQ , Tang YF, et al. D ifferentiation of putative hepatic stem cells derived from adult rats into mature hepatocytes in the p resence of ep idermal grow th facto r and hepatocyte growth facto r. D ifferentiation, 2003, 71 (425): 281-290.
[47] Yamada T, Yo sh ikawaM , Kanda S, et al. In v itro differentiation of embryonic stem cells into hepatocyte21ike cells identified by cellular up take of indocyanine green. Stem Cells, 2002, 20 ( 2):146-154
[48] Peterson BE, Bowen WC, Patrene KD, et al. Bone marrow as a po tential source of hepatic source of hepatic oval cells. Science, 1999, 284 (5417): 1168-1170.
[49] Theise ND, Badve S, Saxcna R, et al. Derivation of hepatocytes from bone marrow cells in m ice radiation induced mycloablation. Hepato logy, 2001, 31 (1) :235-240.
[50] Shen CN , Slack JM , To sh D. Mo lecullar basis of transdifferentiation of pancrease to liver. N at Cell Bio 1, 2000, 2 (12): 879-887.
[51] Krakow sk i ML , Kritzik MR, Jones EM , et al. Pancreatic expression of keratinocyte grow th facto r leads to differentiation ofislet hepatocytes and p ro liferation of duct cells. Am J Patho 1, 1999, 154 (3): 683-691
[52] TimothyW. Austin Eric Lagasse. Hepatic regeneration from hemato2 poietic stem cells[ J ]. M echanism s of Developm ent, 2003, 120: 131 -135.
[53] 黄淑帧,作显辉,彭智增,等1例移植人造血干细胞的山头组织中人特异蛋白的表达[J]1中华医学杂志,2002,82:894-898.
[54] Clarke DL, Jonansson CB,Wilbertz J, et al. Generalized potential of adult neural stem cells[ J ]. Science, 2000, 288: 1660 - 1663.
[55] Kenji Okumura, Kimitoshi Nakamura, Yuichiro Hisatomi, et al. Sali2 vary gland p rogenitor cells induced by duct ligation differentiate into hepatic and pancreatic lineages [ J ]. Hepatology, 2003, 38: 104 -113.
[56] Takashima S; Ise H; Zhao P; Human amniotic epithelial cells possess hepatocyte-like characteristics and functions. Cell Struct Funct. 2004; 29(3):73-84.
[57] Miki T; Lehmann T; Cai H; et al. Stem cell characteristics of amniotic epithelial cells. Stem Cells. 2005; 23(10): 1549-59.
[58] Ilancheran S; Michalska A; Peh G; Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9 [Epub ahead of print].
[59] Lannoy V J a. JBiolChem, 2000, 275(29): 22098-22103.
[60] Llbbrecht L, DesmetV, Van Damme B, et al. Deep intralobular extension of Human hapatic 'progenitorcells'correlates with parenchymal inflammation in chronic viral hepatitis: can 'progenitor cells' migrate? [J]. J Pathol. 2000,192(3):373-378.
[61] Libbrecht L, Desmet V, Van Damme B, et al. The immunohistochemical Phenotype of dysplastic foci in human liver: corelation with putative progenitor cells[J]. J Hepatol. 2000, 33(1):76-84.
[62] Miyazaki M, Akiyama I, Sakaguchi M, et al. Improved conditions to induce Hepatocytes from rat bone marrow cells in culture[J]. Biochem Biophys Res Commun.2002, 298(1):24-30.
[63] Yoshida K, Seto-Ohshima A, and sinohara H. Sequcncing of cDNA encoding Serum albumin and its extrahepatic synthesis in the Mongolian gerbil Meirones unguiculatus[J]. DNARes.1997, 4(5):351-360.
[64] Dodson C S, Rengarajan K, Gewant H D, et al. Extra-hepatic expression of Serum albumin mRNA in mosue retina[J]. Crue Eye Res.2001, 22(3): 182-189
[65] Adler a J and Edwards R B. Human interphotoreceptor matrix contain serum albumin and retinol-bindnig proteinl[J]. Exp Eye Res. 2000, 70(2):227-234.
[66] Nalon J L, Tratner I, Poliard A, et al. Albumin and alpha-fetoprotein gene expression in various nonhepatic rat tissues [J]. J Biol Chem. 1988, 263(23):11436-11542.
[67] Jun Ping Wei, Tian Shu Zhang, Shigeyuki Kawa, Toru Aizawa, Masao Ota,Toshihiro Akaike, Kiyoshi Kato, Ikuo Konishi, and Toshi Nikaido, et al. Human Amnion-Isolated Cells Normalize Blood Glucose in Streptozotocin-Induced Diabetic Mice. Cell Transplantation. VOL. 12. Pp. 2003, 545-552.
[68] Sivakami Hancheran, Anna Michalska, Gary Peh,, et al. Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential.
[69] 杨明智,陈积圣.永生化肝细胞移植.国外医学外科学分册,2001,28(5):257-259
[70] Guha C ,Chowdhury NR,C howdhury JR,et al.Reversibly immortalized human hepatcytes:an eternal fountain of liver support?Hepatology,2000,32 :440-44.
[71] 阂军,陈积圣.肝细胞移植研究进展.第四届中华实验外科学术会议报告。2002年5月,南京
[72] Gupta S .Hepatocyte transplantation:emerging insights into mechanism of liver repopulation and their relevance to potential therapies.J Hepatology,l 999,30 :162-170.
[73]张杰,陈积圣.肝细胞移植研究现状.国外医学外科学分册,2000,27:196-198.
[74]王坤,陈积圣.肝细胞移植中的几个关键问题.国外医学外科学分册,1998,25:130-132.
[75]Song E, Chen JS, Min J, et al. FasL expression of splenocytes and its correlation with hepatocytic apotosis during intrasplenicaliy allogeneic hepatocyte transplantation.Asia J Surgery,2000,23(2):163-168.
[76]M.L.Dumble,E.J.Croager,G.C.T.Yeoh,et al.Generation and characterization of p53 null transforemed hepatic progenitor cell:oval cells give rise tohepatocellular carcinoma.Carcinogenesis,2002,23,3:435-445.
[77]Haochuan Li, 1 Jerry Y. Niederkorn,et al. Immunosuppressive Factors in Amniotic Epithelial Cells. IOVS, 2005,46:900-907
[78]Cai J , Ito M , N agata H, et al. Treatment of liver failure in rats w ith end stage cirrhosis by t ransp lantat ion of immo rtalized hepatocytes.Hepatology, 2002; 36: 386
[79]Gupta S , Rajvansh i P, Lee CD. Integration of transplanted hepatocytes into host liver plates demonstrated with dipeptidyl peptidase Ⅳ-deficientrats. Proc Natl Acad SciUSA, 1995; 92: 5860
[80]S.Y. Onga, H. Daia, K. W. Leong,et al, Inducing hepatic differentiation of human mesenchymal stem cells in pellet culture.Biomaterials, 2006,27:4087-4097
[81]I. Tabei, H. Hashimoto, I. Ishiwata, Characteristics of Hepatocytes Derived From Early ES Cells and Treatment of Surgically Induced Liver Failure Rats by Transplantation. Transplantation Proceedings,2005,37:262-264.
[82]Chinzei R ,Tanaka Y ,Shimizu-Saito K ,et al. Embryoid-body cells derived from a mouse embryonic stem cell line show diferentiation into functional hepatocytes.Hepatology,2002,3 6(1):22-29
[83]Choi D ,Oh HJ,Chang UJ,et al .In vivo differentiation of mouse embryonic stem cells into hepatocytes. Cell Transplant,20 02,11(4):359-68
[84]Rambhatla L ,Chiu C P, Kundu P .Generation of hepatocyte-like cells from human embryonic stem cells. Cell Transplant,20 03,12 (1):1-1
[85]展玉涛；魏来；陈红松等,骨髓干细胞在大鼠肝纤维化形成环境中的分化.中华肝脏病杂志,2003,11:673-675
[86]郭菲菲,刘晓萍,高胜利等.骨髓基质干细胞向肝细胞的定向分化及体内移植.世界华人消化杂志,2007,15(5):464-4707
[87]Wang P , Wang J H., Yan Z P, et al. Expression of hepatocyte-like phenotypes in bone marrow stromal cells after HGF inductions. Biochem Biophys Res Commun ,2004, 320(3):712-716.
[88] Lee KD, Kuo TK, Whang-Peng J, et al. In vitro hepatic differentiation of human mesenchymal stem cells.Hepatology. 2004 , 40(6): 1275-84.
[89]Yamamoto N, Terai S, Ohata S, et al. A subpopulation of bone cells depleted by a noval antibody,anti-Live8,is useful for cell therapy to repair damaged liver. Biochem Biophys Res Commun ,2004,313:1110-1118.
[90]Sato Y, Araki H, Kato J, et al. Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion.Blood.2005, 106(2):756-63.
[91]Dabeva MD, Petkov PM, Sandhu J, et al. Proliferation and differentiation of fetal liver epithelial progenitor cells after transplantation into adult rat liver. Am J Pathol.2000, 156(6):2017-31.
[92]Kollet O, Shivtiel S, Chen YQ, et al. HGF, SDF-1,and MMP-9 are involved in stress-induced human CD34+ stem cell recruitment to the liver, J. Clin. Invest. 2003,112:160-169.
[93]Hatch HM, Zheng D, Jorgensen ML, Petersen BE, SDF-lalpha/CXCR4: a mechanism for hepatic oval cell activation and bone marrow stem cell recruitment to the injured liver of rats, Cloning Stem Cells 2002, 4 : 339-351.
[94]丹尼尔R.马沙克,理查德L.甘德,大卫·戈特利布主编.干细胞生物学[M].第一版.刘景生等译.北京:化学工业出版社,2004,331-339.Marshak DR,.Gardner RL, Gottlieb D. Stem cell biology.The first edition,chemical industrial press,Bejing,2004; 331-339
[95]Hall P A, Watt F M. Stem cells:the generation and maintenance of cellular diversity[J]. Development, 1989,106: 619-633.
[96]Quesenbry P J, Becker P S. Stem cell homing: Rolling,crawling,and nesting[J]. Proc. Natl. Acad. Sci, 1998,95: 15155-15157.
[97]Takeda M,Yamamoto M, Isoda K, Higashiyama S, Hirose M, Ohgushi H, Kawase M, Yagi K. Availability of bone marrow stromal cells in three dimensional coculture with hepatocytes and transplantation into liver-damaged mice. J Biosci Bioeng 2005; 100:77-81
[98]Suzuki A, Iwama A, Miyashita H, Nakauchi H, Taniguchi H. Role for growth factors and extracellular matrix in controlling differentiation of prospectively isolated hepatic stem cells. Development 2003; 130: 2513-2524
[99]Dumble ML, Croager EJ, Yeoh GC, Quail EA. Generation and characterization of p53 null transformed hepatic progenitor cells: oval cells give rise to hepatocellular carcinoma. Carcinogenesis 2002; 23: 435-445
[100]Xiao JC. Small epithelial cells in human liver cirrhosis exhibit features of hepatic stem like cells : immunohistochemical , electron microscopic and immunoelectron microscopic findings [ J ] .Histopathology , 2003 ,42 (2): 141-149.
[101] Shiga ,A ,Shirota ,K. Vimentin/ cytokeratin coexpression foci in a well differentiated canin hepatocellular carcinoma[J ] . J Vet Med Sci , 2000 ,62 (2) : 199-202.
[102]Chinzei R, Tanaka Y, Shimizu-Saito K,. Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes. Hepatology 2002,36: 22-29.
[103]T Miki,T Lehmann,H B Cai,et al.Stem Cell Characteristics of Amniotic Epithelial Cells,Stem Cells 2005;23:1549-1559.
[104]Akle CA, Adinolfi M, Welsh KI et al. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers. Lancet 1981;2:1003-1005.
[105]Scaggiante B, Pineschi A, Sustersich M et al. Successful therapy of Niemann-Pick disease By Implantation of human amniotic membrane. Transplantation,1987,44:59-61.
[106]Tylki-Szymanska A, Maciejko D, Kidawa M et al. Amniotic tissue transplantation as a trial of treatment in some lysosomal storage diseases.J Inherit MetabDis 1985,8:101-104.
[107] Yeager AM, Singer HS, Buck JR et al. A therapeutic trial of amniotic epithelial cell implantation in patients with lysosomal storage diseases. Am J Med Genet 1985,22:347-355.
[108]Sakuragawa N, Yoshikawa H, Sasaki M. Amniotic tissue transplantation:clinical and biochemical evaluations for some lysosomal storage diseases. Brain Dev1992,14:7-11.
1. Travis, John. A Stimulating New Approach To Cancer Treatment. Science; 1993,259; (5093): 310-311.
2. Petersen BE, Hepatic"Stem"Cells: Coming Full Circle [ J ]. B lood cells; M olecules and diaeases; 2001; 27: 590 - 600.
3. TimothyW,Austin Eric Lagasse. Hepatic regeneration from hematopoietic stem cells. M echanism s of Developm ent; 2003; 120: 131 -135.
4. Wang X,Ai-DhalimyM,Lagasse E, et al. Liver repopulation and correctin ofmetabolic liver disease by transp lanted adultmouse pancreat2ic cells[ J ]. Am J Pathol; 2001; 158:571-579.
5. Clarke DL, Jonansson CB,Wilbertz J, et al. Generalized potential of adult neural stem cells[ J ]. Science; 2000; 288:1660-1663.5; 7
6. Kenji Okumura, Kimitoshi Nakamura, Yuichiro Hisatomi, et al. Salivary gland p rogenitor cells induced by duct ligation differentiate intohepatic and pancreatic lineages [ J ]. Hepatology; 2003; 38:104-113.
7. Ilancheran S, Michalska A, Peh G, Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9, EPUB.
8. Y Haruna, K Saito, S Spaulding,et al. Identification of bipotential progenitor cells in human liver development; Hepatology; 1996; 23:476-481.
9. Faris RA, Konkin T, Halpert G. Liver stem cells: a potential source of hepatocytes for the treatment of human liver disease. A rtif O rgans; 2002; 25:513-521.
10. Tanimizu N,Nishikawa M,Saito H, Tsujimura T,Miyajima A. Isolation of hepatoblasts based on the expression of Dlk/Pref-1. J Cell Sci. 2003; 116(Pt 9):1775-86.
ll.Petersen BE,Goff JP,Greenberger JS,Michalopoulos GK.Hepatic oval cells express the h ematopoietic stem cell markerThy-1 in the rat.hepatology,1998;27(2):433-445.
12. Petersen BE.Hepatic stem cell:coming full circle , Blood cells. Mol Dis ,2001:27:590-600.
13. Grisham JW: Migration of hepatocytes along hepatic plates and stem cell-fed hepatocyte lineage. Am J Pathol 1994; 144:849-854.
14. Markus Grompe. Pancreatic hepatic switches in vivo. Mechanism s of Development; 2003; 120:99-106.
15. Oh SH, Witek RP, Bae SH, Bone marrow-derived hepatic oval cells differentiate into hepatocytes in 2-acetylaminofluorene/partial hepatectomy-induced liver regeneration.Gastroenterology. 2007; 132(3): 1077-87.
16. KYM N, EMMA J, JOHN K, et al. Oval cell-mediated liver regeneration: Role of cytokines and growth factors. J of Gastroenterol and Hepatol 2003, 18: 4-12
17. Tsuchiya A,Heike T, Baba S,Long-term culture of postnatal mouse hepatic stem/progenitor cells and their relative developmental hierarchy. Stem Cells. 2007; 25(4):895-902.
18. Leite AR, Correa-Giannella ML, Dagli ML, Fibronectin and laminin induce expression of islet cell markers in hepatic oval cells in culture. Cell Tissue Res. 2007 Mar; 327(3):529-37.
19. Yovchev MI, Grozdanov PN, Joseph B, Novel hepatic progenitor cell surface markers in the adult rat liver. Hepatology. 2007; 45(1):139-49.
20. Coleman W,Grisham J.Epithelial-like stem cell of the rodent liver.In:Strain AJ,Diehl A,eds. Liver growth and repair.London:Chapman and Hall,1998:50-99.
21. Crosby HA,Kelly DA,et al.Human hepatic stem-like cells isolated usingc-kit or CD34 can differentiate into biliary epithelium.Gastroenterology 2001,120: 534- 544.
22. Sandhu JS, Petokov PM, et al. Stem cell properties and repopulation of the rat liver by fetal liver epithelial progenitor cells. Am J Pathol 2001,159 (4):1323-1334.
23. LazaroCA,RhimJ A,et al.Generation of hepatocyte from oval cell precursors in culture. Cancer Res.l998,58:5514-5522.
24. Petersen BE.Hepatic stem cellsxoming full circle.Blood Cells,Molecules,and Diseases2001,27(3):590-600.
25. Mitaka T,Mikami M,Sattler GL,Pitot HC,Mochizuki Y.Small cell colonies appearin the primary culture of adult rat hepatocytes in the presence of nicotinamide and epidermal growth factor. Hepatology, 1992; 16:440-447
26. Mitaka T, Sato F, Mizuguchi T, Yokono T, Mochizuki Y.Reconstruction of hepatic organoid by rats mall hepatocytes and hepatic nonparenchymal cells.hepatology . 1999;29:115-125
27. LemireJ M,Shiojiri N,Fausto N.Oval cell proliferation and the origin of small hepatocytes in liver injury induced by D-galactosamine.Am J Pathol 1991;1393:535-552
28. Herrera MB, Bruno S, Buttiglieri S, Isolation and characterization of a stem cell population from adult human liver. Stem Cells. 2006 Dec; 24(12):2840-50.
29. Mitaka T, MikamiM,Sattler GL, et al. Small cell colonies appear in the p rimary culture of adult rat hap tocytes in the p resence of nicotinamide and ep idermal growth factor[ J ]. Hepatology; 1992; 16: 440-4471.
30. Gordon G, ColemanW,Grisham J, et al. Temporal analysis of hepatocyte differentiation by small hepatocyte-like p rogenitor cells during liver regeneration in retrorsine exposed rats. Am J Pathol; 2000; 157:771-7861.
31. Sidler Pfandler MA, Hochli M, Inderbitzin D, Small hepatocytes in culture develop polarized transporter expression and differentiation. J Cell Sci. 2004; 117(Pt18):4077-87.
32. Badve S, Logdberg L,Lal A,Small cells in hepatoblastoma lack "oval" cell phenotype.Mod Pathol. 2003; 16(9):930-6.
33. Dahlke MH, Popp FC, Bahlmann FH, Liver regeneration in a retrorsine/CC14-induced acute liver failure model: do bone marrow-derived cells contribute? J Hepatol. 2003 Sep; 39(3):365-73.
34. Chinzer R, Tanaka Y,Shimizu Saito K,et al. Embryoid2body cells derived from a mouse embryonic stem cells line show differentiation into functional hepatocytes. Hepatology; 2002; 36:22-29.
35. Yamada T,Yoshikawa M, Kanda S, et al. In vitro diffeentiation of embryonic stem cells in hepatocyte-like cell identified by celllar up take of indecyanine green. Stem Cells; 2002; 20:146-154.
36. Hamazaki T,Liboshi YOka M, et al. Hepatic maturation in differentiating embryonic stem cells in vitro. FEBS Lett; 2001; 497: 15-19.
37. Ek M, Soderdahl T, Kuppers-Munther B, Expression of drug metabolizing enzymes in hepatocyte-like cells derived from human embryonic stem cells.Biochem Pharmacol. 2007; 74 (3): 496-503.
38. Hay DC, Zhao D, Ross A, Direct differentiation of human embryonic stem cells to hepatocyte-like cells exhibiting functional activities. Cloning Stem Cells. 2007; 9(1):51-62.
39. Cai J, Zhao Y,Liu Y, Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology. 2007; 45(5): 1229-39.
40. DANFORTH D N ,HULL R W. The Microscopic Anatomy of the Fetal Membranes wit h Particular Reference to t he Detailed Structure of the Amnion[J] . Am J Obstet Gynecol ,1958 ,75 :536 - 550.
41. BOURNE G L. The Microscopic Anatomy of t he Human Amnion and Chorion[J ] . Am J Obstet Gynecol, 1960 ,79 :1070 -1073.
42. POLLARD S M ,AUE N N ,SYMONDS E M. Scanning Electron Microscope Appearances of Normal human Amnion and Umbilical Cord at Term[ J ].Br J Obstet Gynaecol ,1976 ,83 :470 - 477.
43. 李莹.羊膜在眼科中应用的医学基础及临床研究新进展[J].黑龙江医药科学,2004,27(6):60-62.
44. Takashima S; Ise H; Zhao P; Human amniotic epithelial cells possess hepatocyte-like characteristics and functions. Cell Struct Funct. 2004 ;29(3):73-84.
45. Miki T; Lehmann T; Cai H; et al. Stem cell characteristics of amniotic epithelial cells. Stem Cells. 2005; 23(10):1549-59.
46. Ilancheran S; Michalska A; Peh G; Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9 [Epub ahead of print]
47. MIKI T, LEHMANN T, CA IH, et al. Stem cell characteristics of amniotic ep ithelial cells[ J ]. Stem Cells, 2005, 23 (10): 1549- 1559.
48. AKLE C A, AD INOLF IM, WELSH K I, et al. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers[ J ]. Lancet, 1981, 2(8254): 1003 -1005.
49. H IDENOR IOKAWA, CA OSAMU OKUDA, HAJ IME ARA I, et al. Amniotic epithelial cells transform into neuron - like cells in the ischemic brain[ J ].Neuroreport, 2001, 12: 4003 - 4007.
50. FL IN IAUX I, V IALLET J P, DHOUA ILLY D, et al. Transformation of amnion epithelium into skin and hair follicles[ J ]. Differentiation, 2004, 72 (9 /10) : 558 -565.
51. ADINOL FI M ,A KL E C A ,MCCOLL I. Expression of HLA Antigens ,Beta-microglobulin and Enzymes by Human Amniotic Epit helial Cells[J] .Nature ,1982 ,295:325-327.
52. HOULIHAN J M ,BIRO P A , HARPER H M. The Human Amnion is a Site of MHC Class Ib Expression:Evidence for the Expression of HLAE and HLAG[J]. J Immunol ,1995 ,154 :5665 - 5674.
53. Sutton L , Gadd M ,Masson D Y, et al . Cells BearingClass Ⅱ MHC Antigens in the Human Placenta and Amnio-chorion[J ]. Immunology ,1986 ,58 :23 - 29.
54. Kubo M,Sonada Y,Muramamatsu,et al.Immunogenicity of Human Amniotic Membrane in Experimental Xeno transplantation[J].InvestOphthalmol VisSci,2001 ,42:1539-1546.
55.徐存栓.肝脏干细胞和胰脏干细胞见:章静波,宗书东,马文丽.主编干细胞[M]北京:中国协和医科大学出版社;2003:163-181.
56.Deutsch G; Jung J; ZhengM; et al. A bipotential p recursor population for pancreas and liver within the embryonic endoderm . Development; 2001; 128:871-881.
57.DabevaMD; Hwang BC; Vas SR; et al.Differentiation of pancreatic epithelial proqenitor cells into hepatocytes following transplantation into rat liver. Proc Natl AcasSci USA; 1997; 94:7356-7361.
58.Burke ZD; Shen CN; Ralphs KL; Characterization of liver function in transdifferentiated hepatocytes. J Cell Physiol. 2006 Jan; 206(1): 147-59.
59.Lardon J; De Breuck S; Rooman I; Plasticity in the adult rat pancreas:transdifferentiation of exocrine to hepatocyte-like cells in primary culture.Hepatology. 2004 Jun; 39(6): 1499-507.
60.Yamada S,Terada K,Ueno Y, Sugiyama T,Seno M, Kojima I. Differentiation of adult hepatic stem-like cells into pancreatic endocrine cells. Cell Transplant.2005; 14(9):647-53.
61.Nakajima-Nagata N; Sakurai T; Mitaka T; In vitro induction of adult hepatic progenitor cells into insulin-producing cells.Biochem Biophys Res Commun.2004 Jun 4; 318(3):625-30.
62.Krakowski ML,Kritzik MR,Jones EM, et al.Pancreatic expression of keratinocytes growth factor lead to differentiation of islet hepatocytes and proliferation of duct cells.Am J Pathology, 1999,154 (3 ) :683-691.
63.Petersen BE,BowenWC, Patreme KD, et al. Bonemarrow as a potential source of hepatic oval cells. Science; 1999; 284: 1168-1170.
64.黄淑帧,作显辉,彭智增,等.移植人造血干细胞的山头组织中人.特异蛋白的表达.中华医学杂志;2002;82:894-898.
65.杨明智,闵军,陈积圣.等.体外诱导大鼠骨髓干细胞向肝细胞横向分化.中华 医学杂志;2003:83:603-604
66.Petersen BE; Bowen WC; Patrene KD; et al.Bone marrow as a potential source of hepatic oval cells.Science; 1999; 284:1168-1170.
67.Theise ND,Nimmakayalu M, Gardner R, et al. Liver from bone marrow in humans. Hepatology; 2000; 32:11-16.
68.Malcolm R, Richard P,Rosemar J, et al.Hepatocytes from nonhepatic adult stem cells. Nature ; 2000; 406:257.
69.杜智; 朱争艳;张金卷等,胎儿骨髓源性亚全能干细胞分离培养及诱导肝细胞分化研究。生物医学工程与临床2007;11(1):63-65.
70.Barry FP. Biology and clinical applications of mesenchymal stem cells.Birth Defects Res C Embryo Today. 2003 Aug; 69(3):250-6.
71.Sato Y,Araki H, Kato J, et al. Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion. Blood. 2005 Jul 15; 106(2):756-63.
72.Ong SY, Dai H, Leong KW. Hepatic differentiation potential of commercially available human mesenchymal stem cells.Tissue Eng. 2006; 12(12):3477-85.
73.何念海,赵文利,王明宇.人胎肝非实质间充质肝细胞诱导分化为类肝细胞,中华肝病杂志,2007:15(3):164-169.
74.Banas A; Teratani T; Yamamoto Y; Tokuhara M; Takeshita F; Quinn G;Okochi H; Ochiya T. Adipose tissue-derived mesenchymal stem cells as a source of human hepatocytes.Hepatology. 2007; 46(l):219-28.
75.Fiegel HC; Lange C; KneserU; et al.Fetal and adult liver stem cells for liver regeneration and tissue engineering. J Cell Mol Med. 2006; 10(3):577-87.
76.Myoung WL ,Jeongeun C, et al. Mesenchymal stem cells from crypreserved human umbilical cord blood. Bioehem Biophys Res Commun, 2004, 320(1):273-278
77.Hong SH,Gang EJ, Jeong JA, et al. In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells. Biochem Biophys Res Commun, 2005.330(4):1153-61
78. Philip N. Newsome U. Human cord blood-derived cells can differentiate into hepatocytes in the mouse . Liver with no evidence of celular fusion. Gastroenterology, 2003.124: 1891-1900.
79. Takahisa F, Tetsure H, Hideaki F, et al. Purification of adult hepatic progenitor cells using green fluorescent protein (GFP)-transgenic mice and fluorescence-activated cell sorting. J Hepatol, 2003, 39: 162-170
80. Mihonitou N, Yoshinori S, Katsutoshi I. Purification of Fetal Mouse Hepatoblasts by Magnetic Beads Coated with Monoclonal Anti-E-Cadherin Antibodies and Their In Vitro Culture. Exp Cell Res, 2002, 279: 330-343
81. Hisaya A,Tetsure H,Hideaki F,et al.Enrichment of Hepatic Progenitor Cells From Adult Mouse Liver. Hepatology, 2003. 37: 1385-1394
82. Atherine A, Luzaro EJ, Croager CM.et al.Establishment,characterization, and long-term maintenance of cultures of human fetal hepatocytes. Hepatology, 2003,38(5): 1095-1106
83. WeberA .Immortalization of hepatic Progenitor cells.Pathol Biol,2004;52(2):93-96
84. Dabeva MD,Petkov PM,Sandhu J, et al. Proliferation and differentiation of fetal liver epithelial progenitor cells after transplantation into adult rat liver. ANJ Pathol 2000:156:2017-2031
85. GermainL ,Noel M,Gourdeau H,et al.Promotion of growth and differentiation rat ductural oval cells in primary culture.Cancer Research.l988;46:368-378
86. Lazaro CA,Rhim JA,Yamada Y ,et al.Generation of hepatocytes from oval precursors in culture . Cancer Reasearch,1998;58:5514 - 5522.
87. Yang L,Li S,Hatch H,et al.Invitrotrans-differentiation of adult hepatic stem cells into pancreatic endocrine hormone -producing cells.Proc Natl Acad Sci USA .2002,99(12):8078 - 8083.
88. Suzuki A,Zheng YwYW,Kaneko S,et al.Clonal identification and characterization of self-renewing pluripotent stem cells in development liver.J Cell Biol.2002; 156(1): 173
89. Okano J,Shiota G,Matsumoto K.Hepatocyte growth factor exerts a proliferative effect on oval cells through the PI3K / AKT signaling pathway. Bioehem Biophys Res Commun,2003,309(2): 298-304
90. Matsusaka S, Tsujimura T, Toyosaka A, et al. Role of c-kit receptor tymsine kinase in development of oval cells in the rat 2-acetylaminofluorene / partial hepatectomy model.Hepatology. 1999,29: 670-676.
91. Knight B, Yeoh GCT, Husk KL'et al. Impaired preneoplatic changes and liver tumour form ation in tumour necrosis factor receptor type I knockout mice.J ExpMed, 2000, 192: 1809 1818
92. Overturf K; Al dhalimy M; OU CN; et al. Serial transplantation reveals the stem cell like regenerative potential of adult mouse hepatocytes. Am J Pathol, 1997, 151(5): 1273-1280.
93. Matsumoto K. Yoshitomih, Rossant J, et al. Liver organ ogenesis promoted by endothelial cells prior to vascular function. Science. 2001; 294(5542): 559-563.
94. Jung J, Zheng M, Goldfarb M, et al. Initiation of mammalian fiver development from endoderm by fibroblast growth factors. Science. 1999 ; 284(5422 :1998-2003.
95. Sund NJ,Ang SL,Sackett SD et al. Hepatocyte Nuclear Factor 3p (Foxa2) Is Dispensable for Maintaining the Differentiated State of the Adult Hepatocyte.Mol Cell Biol. 2000 July; 20(14): 5175-5183.
96. Zaret KS. Hepatocyte differentiation: from the endoderm and beyond. Curr Opin Genet Dev, 2001; 11(5): 568-574.
97. Eferl R, Sibilia M, Hilberg F, et al. Functions of c-Jun in fiver and heart development. J CeII Bio. 1999. 145 (51): 1049-1061.
98. Ying Zhang; Xue-Fan Bai; Chang-Xing Huang. Hepatic stem cells: existence and origin. World J Gastroenterol 2003; 9(2):201-204.
99. OmaryMB,Ku No,Toivoal DM, et al. Keratins: guardians of the liver. Hepatology; 2002; 35 (2): 251-256.
100. Heather AC,Stefan H,Luca F,et al. immunolocalization of putative human Liver progenitor cells in livers from patients with end-stage primary biliary cirrhosis and slerosing cholangitis using the monoclonal antibody OV-6.A m JP athol,1998,152:771-779.
101.Orla MC,Maura R,Gerry M, et al.In vitro evidence for the presence of hematopoietic stem cell in adult human liver.Hepatology,1999,29:1193-1198 .
102.Lucy GM,Michael PC,Nlamh N,et al.Differential express of lymphoid and myeloid markers on differenting hematopoietic stem cell in normal and tumor-bearing adult human liver.Hepatology,2000,31:1251-1256.
103.Ulrich B,Heather AC,Pamila R,et al.Expression of the stem cell factor receptor c-kit in normal and diseased pediatric liver:identification of a human hepatic progenitor cell?Hepatology, 1999,30:112-117.
104.Tan Y,Hughes D, Wang X, Costa RH. Adenovirus-mediated increase in HNF-3beta or HNF-3alpha shows differences in levels of liver glycogen and gene expression.Hepatology. 2002 Jan; 35(l):30-9.
105.Hughes DE; StolzDB; YuS; Tan Y; ReddyJK; WatkinsSC; Diehl AM; Costa RH. Elevated hepatocyte levels of the Forkhead box A2 (HNF-3beta) transcription factor cause postnatal steatosis and mitochondrial damage.Hepatology.2003 Jun; 37(6):1414-24.
106.Eleswarapu S,Jiang H.Growth hormone regulates the expression of hepatocyte nuclear factor-3 gamma and other liver-enriched transcription factors in the bovine liver.J Endocrinol. 2005 Jan; 184(l):95-105.
107.Westmacott A; Burke ZD; Oliver G; et al. C/EBPalpha and C/EBPbeta are markers of early liver development.Int J Dev Biol. 2006; 50(7):653-7.
108.Dontu G,A1 - Hajj M; Abdallah WM; et al.Stem cells in normal breast development and breast cancer. Cell Prolif; 2003:59- 72.
109.肖家诚;朱延波;朱上林;等。肝细胞肝癌中卵圆细胞的组织学与超微结构研究.临床与实验病理学杂志;2000;(16):177-179.
110.肖家诚,王天翔,Ruck P等。肝硬化组织中卵圆细胞的光镜、电镜与免疫电镜观察,J Clin Exp Pathol 2002;(1):57-60。
111.Shupe T; Petersen BE. Evidence regarding a stem cell origin of hepatocellular carcinoma.Stem Cell Rev. 2005; 1(3):261-4.
112. Alison MR. Liver stem cells: implications for hepatocarcinogenesis.Stem Cell Rev. 2005; 1(3):253-60.
113. Wu XZ,Yu XH. Bone marrow cells: the source of hepatocellular carcinoma? Med Hypotheses. 2007; 69(1):36-42.
114. Sell S, Pierce G B.Maturation arrest of stem cell differentiation is a common pathyway for the Cellular origin of teratocarcinomas and epithelial cancers.Lablnvest, 1994,70 (1):6-22
115. Hixson DC,Chapman L.McBride A,et al.Antigenic phenotypes common to rat oval cells,primary hepatocellular carcinoma sand developing bile ducts.Carcinogenesis, 1997,18 (6): 1169-1175
116.Sell S. Heterogeneity and plasticity of hepatocyte lineage cells.Hepatology,2001,33(3):738-750
117. Dumble M L,Croager E J, Yeoh G C T, et al.Generation and characterization of p53 null transformed hepatic progenitor cells:oval cells give rise to hepatocellular carcinoma.Carcinogenesis,2002,23(3):435-445
118. Bucher N L R, Farmer S R. Liver regeneration following partial hepatectomy: Genes And Metabolism.In: Stain A,Deihl AM(ed).Liver Growth And Repair.Chapman&Hall:London,l 998,1 -27
119. Thorgeirsson S S .Hepatic stem cells in liver regeneration.FASEBJ, 1996,10 (I1):1249-1256
120. Libbrecht L, Desmst Van Damme B, et al. Deep intralobular extension of human hepatic progenitor cells'correlates with parenchymal inflammation in chronic viral hepatitisxan progenit or cells'migrate?J Pathol,2000,15 6(3):1599-1612
121. Crosby H A, Hubscher S, Fabris L.et al. Immunolocalisation of putative human liver progenitor cells in liver of patients with end-stage primary biliary cirrhosis and sclerosing cholangitis using the antibody OV-6.A mJ P athol,l 998,152(3):771-779
122. Libbrecht L, Desmst V, Van Damme B, et al. Deep intralobular extention of human hepatic progenitor cells'crrelates with parenchymal inflammation in chronic viral hepatitisxan progenitor cells'migrate? J P athol,2000,192(3):373-378
123. Libbrecht L,De Vos R,Cassiman D,et al.Progenitor cells in hepatocellular adenomas.Am J Surg Pathol,2001,25(lt) :1388-1396
124. Lowes KN,Brennan B A ,Yeoh GC,et al.Oval cell numbers in human chronic disease are directly related to diseases severity.A m JP athol,19 99,15 4(2):537-541
125. Gong J Q, Fang C H. Relationship between the oval cells and development of hepatocellular carcinoma in rats.World Chin J Digestol,2002,10(10):1133-1139
126. Zulewski H,AbrahamE J,Gerlach M J,et al.Multipotential nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine,exocrine,and hepatic phenotypes.Diabetes,2001,50(3):521-533
127. WeissmanI L.Stem cells: units of development,units of regeneration,and units inevolution.Cell,2000,100(1):157-168
128 . Norris S,Collins C,Doherty D, et al.Resident heparie lymphocyres are phenotypically different from circulating lymphocytes. Hepatology,1998, 28:84-90.
129. Ruck-P,Xiao-J,Kaiserling-E.Small Epithlial cells and the histogenesis of hepatoblastoma.M ectron microscopic immunoelectron microscopic and immunohistochemical fingings.AmJ Pathol,1996,148:321-329.
130. Robrechts C,Devos R,Scoazec J,et al.Primary liver tumour of intermediate (hepatocyte-bileductcell)phenotype: a progenitor cell tumour?Liver, 1998,18:288 -293.
131. Kazushi S,Katsuya A,Takeshi I,et al .Expression of functional CD40 in hum anh epatocellularca rcinoma.Hepatology, 1999,30:920-926
132. A.M. James S; Camillo R; Bernhard J. International trial of the Edmonton protocol for islet transplantation.N Eng J Med; 2006; 355:1318-1330; 133. Perin EC; DohmannHF; Borojevic R; et al. Transendocardial; autologous bone marrow cell transplantation for severe; chronic ischemic heart failure . Circulation; 2003; 107 (18): 2294-2302.
134. Kumar KS; LefkowitchJ; RussoMW; et al. Successful sequential liver and stem cell transp lantation for hepatic due to p rimaryAlamyloidosis. Gastroenterology; 2003; 122 : 2026 - 2031.
135.姚鹏,胡大荣,王帅,等。自体骨髓干细胞移植治疗慢性重症肝病60例.实用医学杂志2005;19:2143-2145。
136.Lee SW,Wang X,Chowdhury NR, Hepatocyte transplantation: state of the art and strategies for overcoming existing hurdles. Ann Hepatol. 2004 Apr-Jun,3(2):48-53.
137.Ohashi K,Yokoyama T, Yamato M,et al. Engineering functional two- and three-dimensional liver systems in vivo using hepatic tissue sheets. Nat Med. 2007 Jul; 13(7):880-885.
[2] Sanchez-Ramos JR. Neural cells derived from adult bone marrow and umbilicalco rdb lood.J N eurosciR es.2002,6 9(6):880-93.
[3] Theise ND, Badve S, Saxena et al. Derivation of hepatocytes from bonem arrowc ellsin m icea fterra diation-inducedm yeloablation.H epatology, 2000,3 l(1):235-240.
[4] Theise N D, N immakayalu M , Gardner R, et al . Liver from bone marrow In humans. Hepatology,2000,32:11-14.
[5] Thalheimcr U. Capre F..Liver transplantation: making the best out of what we Have Dig Dis Sci,2002,47(5):945-953.
[6] Stockrnanl HB, Ijxermans JN.Prospects for the treatment of acute liver Failure. Eur J Gastroenterol Hepatol,2002,14(2):195-203.
[7] Fox IJ,Chowdhury JR,Kaufman SS,et al. Treatment of the Crigler-Najjar syndrome type 1 with hepatocyte transplantation.N Engl J Med. 1998;338(20): 1422-1426.
[8] Lorenti AS. Hepatic stem cells. Medicina (B Aires).2001;61(sptl):614-620.
[9] Kobayashi N Fuliwara T, Wcsterman KA. et al. Prevention of acute liver failure in rats with reversibly immortalized human hepatocytes. Science. 2000.287(19): 1258-1262.
[10] Cogle CR ,Yachnis AT,Laywell ED, et al. Bone marrow transdifferentiation in brain after transplantation: a retrospective study. Lancet, 2004, 363 : 1432-1437.
[11] Schwartz RE, Reyes M, Koodie L, et al. Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. J Clin Invest, 2002, 109: 1291-1302.
[12] Spees JL, Olson SD, Ylostalo J, et al. Differentiation, cell fusion, and nuclear fusion during ex vivo repair of epithelium by human adult stem cells from bone marrow stroma. Proc Natl Acad Sci USA, 2003, 100: 2397.
[13] 王 萍。 中华肝脏病杂志, 2004, 1 2 : 718
[14] SakuragawaN ,T ohyamaJ ,Y amamotoH . Immunostaining of human amniotic epithelil cells:possible use as a transgenec arrier in gene therapy for in bonrerrors of metabolism.Cell Transplant. 1995;4(3):343-346.
[15] Hunt JS, W ood GW. Interferon-gamma induces class I HLA and beta 2-microg lobulin expression by human amnion cells.J I mmunol.l 986;136(2):364- 367.
[16] Tabei I, Hashimoto H , Ishiwata I, et al . Characteristics of hepatocytes derived from early ES cells and treatment of surgically induced liver failure rats by transplantation. Transplant Proc , 2005 , 37 (1) :262-264.
[17] Nishitai R , Koch CA , Ogata K, et al . Toward the survival and function of xenogeneic hepatocyte grafts. Liver Transpl ,2005 , 11 (1) :39 - 50.
[18] Gunsalus JR,Brady DA,Coulter SM, et al . Reduction of serum cholesterol in Watanabe rabbits by xenogeneic hepatocellular transplantation. Nat Med , 1997 , 3(1) :48-53.
[19] Nagata H , Ito M, Cai J , et al. Treatment of cirrhosis and liver failure in rats by hepatocyte xenotransplantation. Gastroenterology, 2003 , 124(2) :422-431.
[20] Maruyama M, Totsugawa T , Kunieda T , et al . Hepatocyte isolation and transplantation in the pig. Cell Transplant, 2003 , 12(6) :593 -598.
[21] Bilir BM, Guinette D , Karrer F , et al . Hepatocyte transplantation in acute liver failure. Liver Transpl, 2000 , 6 (1) :32-40.
[22] Fisher RA , Bu D , Thompson M, et al . Defining hepatocellular chimerism in a liver failure patient bridged with hepatocyte infusion. Transplantation , 2000 , 69 (2):303-307.
[23] Strom SC , Fisher RA , Thompson M, et al . Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure. Transplantation , 1997,63 (4) :559-569.
[24] Nakamura J , Okamoto T , Schumacher IK, et al . Treatment of surgically induced acute liver failure by transplantation of conditionally immortalized hepatocytes. Transplantation , 1997 , 63(11):1541 - 1547.
[25] Cai J , Ito M, Nagata H , et al. Treatment of liver failure in rats with end-stage cirrhosis by transplantation of immortalized hepatocytes. Hepatology , 2002 , 36(2) :386 -394.
[26] Koyano S, Fukui A, Uchida S, Yamada K, et al. Synthesis and release of activin and noggin by cultured human amniotic epithelial cells. Dev Growth Differ. 2002; 44(2): 103-112.
[27] Sakuragawa N, Misawa H, Ohsugi K, et al. Evidence for active acetylcholine metabolism in human amniotic epithelial cells: applicable to intracerebral allografting for neurologic disease. Neurosci Lett. 1997; 232(l):53-56.
[28] Sankar V, Muthusamy R. Role of human amniotic epithelial cell transplantation in spinal cord injury repair research. Neuroscience. 2003; 118(1):11-17.
[29] Tseng SC, Meller D, Anderson DF, et al. Ex vivo preservation and expansion of human limbal epithelial stem cells on amniotic membrane for treating corneal diseases with total limbal stem cell deficiency. Adv Exp Med Biol. 2002; 506:1323-1334.
[30] akuragawa N, Tohyama J, Yamamoto H. Immunostaining of human amniotic epithelial cells: possible use as a transgene carrier in gene therapy for inborn errors of metabolism. Cell Transplant. 1995; 4(3):343-346.
[31] Sakuragawa N, Enosawa S, Ishii T, et al. Human amniotic epithelial cells are promising transgene carriers for allogeneic cell transplantation into liver. J Hum Genet. 2000; 45 (3): 171-176.
[32] Bhattacharya B, Miura T, Brandenberger R, et al. Gene expression in human embryonic stem cell lines: unique molecular signature. Blood. 2004; 103 (8): 2956-64.
[33] Chambers I, Colby D, Robertson M, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 2003; 113 (5): 643-655.
[34] Cavaleri F, Scholer HR. Nanog: a new recruit to the embryonic stem cell orchestra. Cell. 2003; 113 (5): 551-552.
[35] Brivanlou AH, Gage FH, Jaenisch R, et al.Stem cells. Setting standards for human embryonic stem cells.Science. 2003; 300(5621):865.
[36] Gelse K, von der Mark K. Cartilage regeneration by gene therapy.Curr Gene Ther,2003; 3: 305-317.
[37] Coffin RS, Thomas SK, Thomas NSB. Pure populations of transduced Primary human cells can be produced using GFP expression herpes virus vectors and flow cytometry. Gene Ther, 1998; 5: 718-722.
[38] Soderqvist H , Imerch G, Kihlark M. Intracellular distribution of an integral nuclear pore membrane protein fused to green fluorescent protein locatization of a targeting domain. Eur J Biochem,1997; 250(3) :808-813.
[39] Kalajzic I, Braut A, Guo D, Jiang X, Kronenberg MS, Mina M, Harris MA, Harris SE, Rowe DW, Dentin matrix protein 1 expression during osteoblastic differentiation, generation of an osteocyte GFP transgene . Bone, 2004; 35(1):74-82.28
[40] Toshio Miki,a Thomas Lehmann,a Hongbo Cai,a Donna B. Stolz,b,c Stephen C. Stroma,cStem Cell Characteristics of Amniotic Epithelial Cells.Stem Cells 2005;23:1549 - 1559.
[41 ] Ilancheran S; Michalska A; Peh G; Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9, EPUB .
[42] 裴雪涛,刘大庆.干细胞. 组织器官重建的理想种子细胞. 中国修复重建外科杂志, 2006, 20 (4): 344-348.
[43] Strick2M archand H, Mo ro san S, Charneau P, et al. Bipo tential mouse embryonic liver stem cell lines contribute to liver regeneration and differentiate as bile ducts and hepatocytes. P roc N atl A cad SciU SA, 2004, 101 (22): 8360-8365.
[44] Zheng YW , Taniguch i H. D iversity of hepatic stem cells in the fetal and adult liver. Sem in L iver D is, 2003, 23 (4): 3372-3480.
[45] Paku S, Schnur J , N agy P, et al. O rigin and structural evo lution of the early p ro liferating oval cells in rat liver. Am J Patho 1, 2001, 158 (4): 1313-1323.
[46] He ZP, TanWQ , Tang YF, et al. D ifferentiation of putative hepatic stem cells derived from adult rats into mature hepatocytes in the p resence of ep idermal grow th facto r and hepatocyte growth facto r. D ifferentiation, 2003, 71 (425): 281-290.
[47] Yamada T, Yo sh ikawaM , Kanda S, et al. In v itro differentiation of embryonic stem cells into hepatocyte21ike cells identified by cellular up take of indocyanine green. Stem Cells, 2002, 20 ( 2):146-154
[48] Peterson BE, Bowen WC, Patrene KD, et al. Bone marrow as a po tential source of hepatic source of hepatic oval cells. Science, 1999, 284 (5417): 1168-1170.
[49] Theise ND, Badve S, Saxcna R, et al. Derivation of hepatocytes from bone marrow cells in m ice radiation induced mycloablation. Hepato logy, 2001, 31 (1) :235-240.
[50] Shen CN , Slack JM , To sh D. Mo lecullar basis of transdifferentiation of pancrease to liver. N at Cell Bio 1, 2000, 2 (12): 879-887.
[51] Krakow sk i ML , Kritzik MR, Jones EM , et al. Pancreatic expression of keratinocyte grow th facto r leads to differentiation ofislet hepatocytes and p ro liferation of duct cells. Am J Patho 1, 1999, 154 (3): 683-691
[52] TimothyW. Austin Eric Lagasse. Hepatic regeneration from hemato2 poietic stem cells[ J ]. M echanism s of Developm ent, 2003, 120: 131 -135.
[53] 黄淑帧,作显辉,彭智增,等1例移植人造血干细胞的山头组织中人特异蛋白的表达[J]1中华医学杂志,2002,82:894-898.
[54] Clarke DL, Jonansson CB,Wilbertz J, et al. Generalized potential of adult neural stem cells[ J ]. Science, 2000, 288: 1660 - 1663.
[55] Kenji Okumura, Kimitoshi Nakamura, Yuichiro Hisatomi, et al. Sali2 vary gland p rogenitor cells induced by duct ligation differentiate into hepatic and pancreatic lineages [ J ]. Hepatology, 2003, 38: 104 -113.
[56] Takashima S; Ise H; Zhao P; Human amniotic epithelial cells possess hepatocyte-like characteristics and functions. Cell Struct Funct. 2004; 29(3):73-84.
[57] Miki T; Lehmann T; Cai H; et al. Stem cell characteristics of amniotic epithelial cells. Stem Cells. 2005; 23(10): 1549-59.
[58] Ilancheran S; Michalska A; Peh G; Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9 [Epub ahead of print].
[59] Lannoy V J a. JBiolChem, 2000, 275(29): 22098-22103.
[60] Llbbrecht L, DesmetV, Van Damme B, et al. Deep intralobular extension of Human hapatic 'progenitorcells'correlates with parenchymal inflammation in chronic viral hepatitis: can 'progenitor cells' migrate? [J]. J Pathol. 2000,192(3):373-378.
[61] Libbrecht L, Desmet V, Van Damme B, et al. The immunohistochemical Phenotype of dysplastic foci in human liver: corelation with putative progenitor cells[J]. J Hepatol. 2000, 33(1):76-84.
[62] Miyazaki M, Akiyama I, Sakaguchi M, et al. Improved conditions to induce Hepatocytes from rat bone marrow cells in culture[J]. Biochem Biophys Res Commun.2002, 298(1):24-30.
[63] Yoshida K, Seto-Ohshima A, and sinohara H. Sequcncing of cDNA encoding Serum albumin and its extrahepatic synthesis in the Mongolian gerbil Meirones unguiculatus[J]. DNARes.1997, 4(5):351-360.
[64] Dodson C S, Rengarajan K, Gewant H D, et al. Extra-hepatic expression of Serum albumin mRNA in mosue retina[J]. Crue Eye Res.2001, 22(3): 182-189
[65] Adler a J and Edwards R B. Human interphotoreceptor matrix contain serum albumin and retinol-bindnig proteinl[J]. Exp Eye Res. 2000, 70(2):227-234.
[66] Nalon J L, Tratner I, Poliard A, et al. Albumin and alpha-fetoprotein gene expression in various nonhepatic rat tissues [J]. J Biol Chem. 1988, 263(23):11436-11542.
[67] Jun Ping Wei, Tian Shu Zhang, Shigeyuki Kawa, Toru Aizawa, Masao Ota,Toshihiro Akaike, Kiyoshi Kato, Ikuo Konishi, and Toshi Nikaido, et al. Human Amnion-Isolated Cells Normalize Blood Glucose in Streptozotocin-Induced Diabetic Mice. Cell Transplantation. VOL. 12. Pp. 2003, 545-552.
[68] Sivakami Hancheran, Anna Michalska, Gary Peh,, et al. Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential.
[69] 杨明智,陈积圣.永生化肝细胞移植.国外医学外科学分册,2001,28(5):257-259
[70] Guha C ,Chowdhury NR,C howdhury JR,et al.Reversibly immortalized human hepatcytes:an eternal fountain of liver support?Hepatology,2000,32 :440-44.
[71] 阂军,陈积圣.肝细胞移植研究进展.第四届中华实验外科学术会议报告。2002年5月,南京
[72] Gupta S .Hepatocyte transplantation:emerging insights into mechanism of liver repopulation and their relevance to potential therapies.J Hepatology,l 999,30 :162-170.
[73]张杰,陈积圣.肝细胞移植研究现状.国外医学外科学分册,2000,27:196-198.
[74]王坤,陈积圣.肝细胞移植中的几个关键问题.国外医学外科学分册,1998,25:130-132.
[75]Song E, Chen JS, Min J, et al. FasL expression of splenocytes and its correlation with hepatocytic apotosis during intrasplenicaliy allogeneic hepatocyte transplantation.Asia J Surgery,2000,23(2):163-168.
[76]M.L.Dumble,E.J.Croager,G.C.T.Yeoh,et al.Generation and characterization of p53 null transforemed hepatic progenitor cell:oval cells give rise tohepatocellular carcinoma.Carcinogenesis,2002,23,3:435-445.
[77]Haochuan Li, 1 Jerry Y. Niederkorn,et al. Immunosuppressive Factors in Amniotic Epithelial Cells. IOVS, 2005,46:900-907
[78]Cai J , Ito M , N agata H, et al. Treatment of liver failure in rats w ith end stage cirrhosis by t ransp lantat ion of immo rtalized hepatocytes.Hepatology, 2002; 36: 386
[79]Gupta S , Rajvansh i P, Lee CD. Integration of transplanted hepatocytes into host liver plates demonstrated with dipeptidyl peptidase Ⅳ-deficientrats. Proc Natl Acad SciUSA, 1995; 92: 5860
[80]S.Y. Onga, H. Daia, K. W. Leong,et al, Inducing hepatic differentiation of human mesenchymal stem cells in pellet culture.Biomaterials, 2006,27:4087-4097
[81]I. Tabei, H. Hashimoto, I. Ishiwata, Characteristics of Hepatocytes Derived From Early ES Cells and Treatment of Surgically Induced Liver Failure Rats by Transplantation. Transplantation Proceedings,2005,37:262-264.
[82]Chinzei R ,Tanaka Y ,Shimizu-Saito K ,et al. Embryoid-body cells derived from a mouse embryonic stem cell line show diferentiation into functional hepatocytes.Hepatology,2002,3 6(1):22-29
[83]Choi D ,Oh HJ,Chang UJ,et al .In vivo differentiation of mouse embryonic stem cells into hepatocytes. Cell Transplant,20 02,11(4):359-68
[84]Rambhatla L ,Chiu C P, Kundu P .Generation of hepatocyte-like cells from human embryonic stem cells. Cell Transplant,20 03,12 (1):1-1
[85]展玉涛；魏来；陈红松等,骨髓干细胞在大鼠肝纤维化形成环境中的分化.中华肝脏病杂志,2003,11:673-675
[86]郭菲菲,刘晓萍,高胜利等.骨髓基质干细胞向肝细胞的定向分化及体内移植.世界华人消化杂志,2007,15(5):464-4707
[87]Wang P , Wang J H., Yan Z P, et al. Expression of hepatocyte-like phenotypes in bone marrow stromal cells after HGF inductions. Biochem Biophys Res Commun ,2004, 320(3):712-716.
[88] Lee KD, Kuo TK, Whang-Peng J, et al. In vitro hepatic differentiation of human mesenchymal stem cells.Hepatology. 2004 , 40(6): 1275-84.
[89]Yamamoto N, Terai S, Ohata S, et al. A subpopulation of bone cells depleted by a noval antibody,anti-Live8,is useful for cell therapy to repair damaged liver. Biochem Biophys Res Commun ,2004,313:1110-1118.
[90]Sato Y, Araki H, Kato J, et al. Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion.Blood.2005, 106(2):756-63.
[91]Dabeva MD, Petkov PM, Sandhu J, et al. Proliferation and differentiation of fetal liver epithelial progenitor cells after transplantation into adult rat liver. Am J Pathol.2000, 156(6):2017-31.
[92]Kollet O, Shivtiel S, Chen YQ, et al. HGF, SDF-1,and MMP-9 are involved in stress-induced human CD34+ stem cell recruitment to the liver, J. Clin. Invest. 2003,112:160-169.
[93]Hatch HM, Zheng D, Jorgensen ML, Petersen BE, SDF-lalpha/CXCR4: a mechanism for hepatic oval cell activation and bone marrow stem cell recruitment to the injured liver of rats, Cloning Stem Cells 2002, 4 : 339-351.
[94]丹尼尔R.马沙克,理查德L.甘德,大卫·戈特利布主编.干细胞生物学[M].第一版.刘景生等译.北京:化学工业出版社,2004,331-339.Marshak DR,.Gardner RL, Gottlieb D. Stem cell biology.The first edition,chemical industrial press,Bejing,2004; 331-339
[95]Hall P A, Watt F M. Stem cells:the generation and maintenance of cellular diversity[J]. Development, 1989,106: 619-633.
[96]Quesenbry P J, Becker P S. Stem cell homing: Rolling,crawling,and nesting[J]. Proc. Natl. Acad. Sci, 1998,95: 15155-15157.
[97]Takeda M,Yamamoto M, Isoda K, Higashiyama S, Hirose M, Ohgushi H, Kawase M, Yagi K. Availability of bone marrow stromal cells in three dimensional coculture with hepatocytes and transplantation into liver-damaged mice. J Biosci Bioeng 2005; 100:77-81
[98]Suzuki A, Iwama A, Miyashita H, Nakauchi H, Taniguchi H. Role for growth factors and extracellular matrix in controlling differentiation of prospectively isolated hepatic stem cells. Development 2003; 130: 2513-2524
[99]Dumble ML, Croager EJ, Yeoh GC, Quail EA. Generation and characterization of p53 null transformed hepatic progenitor cells: oval cells give rise to hepatocellular carcinoma. Carcinogenesis 2002; 23: 435-445
[100]Xiao JC. Small epithelial cells in human liver cirrhosis exhibit features of hepatic stem like cells : immunohistochemical , electron microscopic and immunoelectron microscopic findings [ J ] .Histopathology , 2003 ,42 (2): 141-149.
[101] Shiga ,A ,Shirota ,K. Vimentin/ cytokeratin coexpression foci in a well differentiated canin hepatocellular carcinoma[J ] . J Vet Med Sci , 2000 ,62 (2) : 199-202.
[102]Chinzei R, Tanaka Y, Shimizu-Saito K,. Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes. Hepatology 2002,36: 22-29.
[103]T Miki,T Lehmann,H B Cai,et al.Stem Cell Characteristics of Amniotic Epithelial Cells,Stem Cells 2005;23:1549-1559.
[104]Akle CA, Adinolfi M, Welsh KI et al. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers. Lancet 1981;2:1003-1005.
[105]Scaggiante B, Pineschi A, Sustersich M et al. Successful therapy of Niemann-Pick disease By Implantation of human amniotic membrane. Transplantation,1987,44:59-61.
[106]Tylki-Szymanska A, Maciejko D, Kidawa M et al. Amniotic tissue transplantation as a trial of treatment in some lysosomal storage diseases.J Inherit MetabDis 1985,8:101-104.
[107] Yeager AM, Singer HS, Buck JR et al. A therapeutic trial of amniotic epithelial cell implantation in patients with lysosomal storage diseases. Am J Med Genet 1985,22:347-355.
[108]Sakuragawa N, Yoshikawa H, Sasaki M. Amniotic tissue transplantation:clinical and biochemical evaluations for some lysosomal storage diseases. Brain Dev1992,14:7-11.
1. Travis, John. A Stimulating New Approach To Cancer Treatment. Science; 1993,259; (5093): 310-311.
2. Petersen BE, Hepatic"Stem"Cells: Coming Full Circle [ J ]. B lood cells; M olecules and diaeases; 2001; 27: 590 - 600.
3. TimothyW,Austin Eric Lagasse. Hepatic regeneration from hematopoietic stem cells. M echanism s of Developm ent; 2003; 120: 131 -135.
4. Wang X,Ai-DhalimyM,Lagasse E, et al. Liver repopulation and correctin ofmetabolic liver disease by transp lanted adultmouse pancreat2ic cells[ J ]. Am J Pathol; 2001; 158:571-579.
5. Clarke DL, Jonansson CB,Wilbertz J, et al. Generalized potential of adult neural stem cells[ J ]. Science; 2000; 288:1660-1663.5; 7
6. Kenji Okumura, Kimitoshi Nakamura, Yuichiro Hisatomi, et al. Salivary gland p rogenitor cells induced by duct ligation differentiate intohepatic and pancreatic lineages [ J ]. Hepatology; 2003; 38:104-113.
7. Ilancheran S, Michalska A, Peh G, Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9, EPUB.
8. Y Haruna, K Saito, S Spaulding,et al. Identification of bipotential progenitor cells in human liver development; Hepatology; 1996; 23:476-481.
9. Faris RA, Konkin T, Halpert G. Liver stem cells: a potential source of hepatocytes for the treatment of human liver disease. A rtif O rgans; 2002; 25:513-521.
10. Tanimizu N,Nishikawa M,Saito H, Tsujimura T,Miyajima A. Isolation of hepatoblasts based on the expression of Dlk/Pref-1. J Cell Sci. 2003; 116(Pt 9):1775-86.
ll.Petersen BE,Goff JP,Greenberger JS,Michalopoulos GK.Hepatic oval cells express the h ematopoietic stem cell markerThy-1 in the rat.hepatology,1998;27(2):433-445.
12. Petersen BE.Hepatic stem cell:coming full circle , Blood cells. Mol Dis ,2001:27:590-600.
13. Grisham JW: Migration of hepatocytes along hepatic plates and stem cell-fed hepatocyte lineage. Am J Pathol 1994; 144:849-854.
14. Markus Grompe. Pancreatic hepatic switches in vivo. Mechanism s of Development; 2003; 120:99-106.
15. Oh SH, Witek RP, Bae SH, Bone marrow-derived hepatic oval cells differentiate into hepatocytes in 2-acetylaminofluorene/partial hepatectomy-induced liver regeneration.Gastroenterology. 2007; 132(3): 1077-87.
16. KYM N, EMMA J, JOHN K, et al. Oval cell-mediated liver regeneration: Role of cytokines and growth factors. J of Gastroenterol and Hepatol 2003, 18: 4-12
17. Tsuchiya A,Heike T, Baba S,Long-term culture of postnatal mouse hepatic stem/progenitor cells and their relative developmental hierarchy. Stem Cells. 2007; 25(4):895-902.
18. Leite AR, Correa-Giannella ML, Dagli ML, Fibronectin and laminin induce expression of islet cell markers in hepatic oval cells in culture. Cell Tissue Res. 2007 Mar; 327(3):529-37.
19. Yovchev MI, Grozdanov PN, Joseph B, Novel hepatic progenitor cell surface markers in the adult rat liver. Hepatology. 2007; 45(1):139-49.
20. Coleman W,Grisham J.Epithelial-like stem cell of the rodent liver.In:Strain AJ,Diehl A,eds. Liver growth and repair.London:Chapman and Hall,1998:50-99.
21. Crosby HA,Kelly DA,et al.Human hepatic stem-like cells isolated usingc-kit or CD34 can differentiate into biliary epithelium.Gastroenterology 2001,120: 534- 544.
22. Sandhu JS, Petokov PM, et al. Stem cell properties and repopulation of the rat liver by fetal liver epithelial progenitor cells. Am J Pathol 2001,159 (4):1323-1334.
23. LazaroCA,RhimJ A,et al.Generation of hepatocyte from oval cell precursors in culture. Cancer Res.l998,58:5514-5522.
24. Petersen BE.Hepatic stem cellsxoming full circle.Blood Cells,Molecules,and Diseases2001,27(3):590-600.
25. Mitaka T,Mikami M,Sattler GL,Pitot HC,Mochizuki Y.Small cell colonies appearin the primary culture of adult rat hepatocytes in the presence of nicotinamide and epidermal growth factor. Hepatology, 1992; 16:440-447
26. Mitaka T, Sato F, Mizuguchi T, Yokono T, Mochizuki Y.Reconstruction of hepatic organoid by rats mall hepatocytes and hepatic nonparenchymal cells.hepatology . 1999;29:115-125
27. LemireJ M,Shiojiri N,Fausto N.Oval cell proliferation and the origin of small hepatocytes in liver injury induced by D-galactosamine.Am J Pathol 1991;1393:535-552
28. Herrera MB, Bruno S, Buttiglieri S, Isolation and characterization of a stem cell population from adult human liver. Stem Cells. 2006 Dec; 24(12):2840-50.
29. Mitaka T, MikamiM,Sattler GL, et al. Small cell colonies appear in the p rimary culture of adult rat hap tocytes in the p resence of nicotinamide and ep idermal growth factor[ J ]. Hepatology; 1992; 16: 440-4471.
30. Gordon G, ColemanW,Grisham J, et al. Temporal analysis of hepatocyte differentiation by small hepatocyte-like p rogenitor cells during liver regeneration in retrorsine exposed rats. Am J Pathol; 2000; 157:771-7861.
31. Sidler Pfandler MA, Hochli M, Inderbitzin D, Small hepatocytes in culture develop polarized transporter expression and differentiation. J Cell Sci. 2004; 117(Pt18):4077-87.
32. Badve S, Logdberg L,Lal A,Small cells in hepatoblastoma lack "oval" cell phenotype.Mod Pathol. 2003; 16(9):930-6.
33. Dahlke MH, Popp FC, Bahlmann FH, Liver regeneration in a retrorsine/CC14-induced acute liver failure model: do bone marrow-derived cells contribute? J Hepatol. 2003 Sep; 39(3):365-73.
34. Chinzer R, Tanaka Y,Shimizu Saito K,et al. Embryoid2body cells derived from a mouse embryonic stem cells line show differentiation into functional hepatocytes. Hepatology; 2002; 36:22-29.
35. Yamada T,Yoshikawa M, Kanda S, et al. In vitro diffeentiation of embryonic stem cells in hepatocyte-like cell identified by celllar up take of indecyanine green. Stem Cells; 2002; 20:146-154.
36. Hamazaki T,Liboshi YOka M, et al. Hepatic maturation in differentiating embryonic stem cells in vitro. FEBS Lett; 2001; 497: 15-19.
37. Ek M, Soderdahl T, Kuppers-Munther B, Expression of drug metabolizing enzymes in hepatocyte-like cells derived from human embryonic stem cells.Biochem Pharmacol. 2007; 74 (3): 496-503.
38. Hay DC, Zhao D, Ross A, Direct differentiation of human embryonic stem cells to hepatocyte-like cells exhibiting functional activities. Cloning Stem Cells. 2007; 9(1):51-62.
39. Cai J, Zhao Y,Liu Y, Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology. 2007; 45(5): 1229-39.
40. DANFORTH D N ,HULL R W. The Microscopic Anatomy of the Fetal Membranes wit h Particular Reference to t he Detailed Structure of the Amnion[J] . Am J Obstet Gynecol ,1958 ,75 :536 - 550.
41. BOURNE G L. The Microscopic Anatomy of t he Human Amnion and Chorion[J ] . Am J Obstet Gynecol, 1960 ,79 :1070 -1073.
42. POLLARD S M ,AUE N N ,SYMONDS E M. Scanning Electron Microscope Appearances of Normal human Amnion and Umbilical Cord at Term[ J ].Br J Obstet Gynaecol ,1976 ,83 :470 - 477.
43. 李莹.羊膜在眼科中应用的医学基础及临床研究新进展[J].黑龙江医药科学,2004,27(6):60-62.
44. Takashima S; Ise H; Zhao P; Human amniotic epithelial cells possess hepatocyte-like characteristics and functions. Cell Struct Funct. 2004 ;29(3):73-84.
45. Miki T; Lehmann T; Cai H; et al. Stem cell characteristics of amniotic epithelial cells. Stem Cells. 2005; 23(10):1549-59.
46. Ilancheran S; Michalska A; Peh G; Stem Cells Derived from Human Fetal Membranes Display Multi-Lineage Differentiation Potential. Biol Reprod. 2007 May 9 [Epub ahead of print]
47. MIKI T, LEHMANN T, CA IH, et al. Stem cell characteristics of amniotic ep ithelial cells[ J ]. Stem Cells, 2005, 23 (10): 1549- 1559.
48. AKLE C A, AD INOLF IM, WELSH K I, et al. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers[ J ]. Lancet, 1981, 2(8254): 1003 -1005.
49. H IDENOR IOKAWA, CA OSAMU OKUDA, HAJ IME ARA I, et al. Amniotic epithelial cells transform into neuron - like cells in the ischemic brain[ J ].Neuroreport, 2001, 12: 4003 - 4007.
50. FL IN IAUX I, V IALLET J P, DHOUA ILLY D, et al. Transformation of amnion epithelium into skin and hair follicles[ J ]. Differentiation, 2004, 72 (9 /10) : 558 -565.
51. ADINOL FI M ,A KL E C A ,MCCOLL I. Expression of HLA Antigens ,Beta-microglobulin and Enzymes by Human Amniotic Epit helial Cells[J] .Nature ,1982 ,295:325-327.
52. HOULIHAN J M ,BIRO P A , HARPER H M. The Human Amnion is a Site of MHC Class Ib Expression:Evidence for the Expression of HLAE and HLAG[J]. J Immunol ,1995 ,154 :5665 - 5674.
53. Sutton L , Gadd M ,Masson D Y, et al . Cells BearingClass Ⅱ MHC Antigens in the Human Placenta and Amnio-chorion[J ]. Immunology ,1986 ,58 :23 - 29.
54. Kubo M,Sonada Y,Muramamatsu,et al.Immunogenicity of Human Amniotic Membrane in Experimental Xeno transplantation[J].InvestOphthalmol VisSci,2001 ,42:1539-1546.
55.徐存栓.肝脏干细胞和胰脏干细胞见:章静波,宗书东,马文丽.主编干细胞[M]北京:中国协和医科大学出版社;2003:163-181.
56.Deutsch G; Jung J; ZhengM; et al. A bipotential p recursor population for pancreas and liver within the embryonic endoderm . Development; 2001; 128:871-881.
57.DabevaMD; Hwang BC; Vas SR; et al.Differentiation of pancreatic epithelial proqenitor cells into hepatocytes following transplantation into rat liver. Proc Natl AcasSci USA; 1997; 94:7356-7361.
58.Burke ZD; Shen CN; Ralphs KL; Characterization of liver function in transdifferentiated hepatocytes. J Cell Physiol. 2006 Jan; 206(1): 147-59.
59.Lardon J; De Breuck S; Rooman I; Plasticity in the adult rat pancreas:transdifferentiation of exocrine to hepatocyte-like cells in primary culture.Hepatology. 2004 Jun; 39(6): 1499-507.
60.Yamada S,Terada K,Ueno Y, Sugiyama T,Seno M, Kojima I. Differentiation of adult hepatic stem-like cells into pancreatic endocrine cells. Cell Transplant.2005; 14(9):647-53.
61.Nakajima-Nagata N; Sakurai T; Mitaka T; In vitro induction of adult hepatic progenitor cells into insulin-producing cells.Biochem Biophys Res Commun.2004 Jun 4; 318(3):625-30.
62.Krakowski ML,Kritzik MR,Jones EM, et al.Pancreatic expression of keratinocytes growth factor lead to differentiation of islet hepatocytes and proliferation of duct cells.Am J Pathology, 1999,154 (3 ) :683-691.
63.Petersen BE,BowenWC, Patreme KD, et al. Bonemarrow as a potential source of hepatic oval cells. Science; 1999; 284: 1168-1170.
64.黄淑帧,作显辉,彭智增,等.移植人造血干细胞的山头组织中人.特异蛋白的表达.中华医学杂志;2002;82:894-898.
65.杨明智,闵军,陈积圣.等.体外诱导大鼠骨髓干细胞向肝细胞横向分化.中华 医学杂志;2003:83:603-604
66.Petersen BE; Bowen WC; Patrene KD; et al.Bone marrow as a potential source of hepatic oval cells.Science; 1999; 284:1168-1170.
67.Theise ND,Nimmakayalu M, Gardner R, et al. Liver from bone marrow in humans. Hepatology; 2000; 32:11-16.
68.Malcolm R, Richard P,Rosemar J, et al.Hepatocytes from nonhepatic adult stem cells. Nature ; 2000; 406:257.
69.杜智; 朱争艳;张金卷等,胎儿骨髓源性亚全能干细胞分离培养及诱导肝细胞分化研究。生物医学工程与临床2007;11(1):63-65.
70.Barry FP. Biology and clinical applications of mesenchymal stem cells.Birth Defects Res C Embryo Today. 2003 Aug; 69(3):250-6.
71.Sato Y,Araki H, Kato J, et al. Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion. Blood. 2005 Jul 15; 106(2):756-63.
72.Ong SY, Dai H, Leong KW. Hepatic differentiation potential of commercially available human mesenchymal stem cells.Tissue Eng. 2006; 12(12):3477-85.
73.何念海,赵文利,王明宇.人胎肝非实质间充质肝细胞诱导分化为类肝细胞,中华肝病杂志,2007:15(3):164-169.
74.Banas A; Teratani T; Yamamoto Y; Tokuhara M; Takeshita F; Quinn G;Okochi H; Ochiya T. Adipose tissue-derived mesenchymal stem cells as a source of human hepatocytes.Hepatology. 2007; 46(l):219-28.
75.Fiegel HC; Lange C; KneserU; et al.Fetal and adult liver stem cells for liver regeneration and tissue engineering. J Cell Mol Med. 2006; 10(3):577-87.
76.Myoung WL ,Jeongeun C, et al. Mesenchymal stem cells from crypreserved human umbilical cord blood. Bioehem Biophys Res Commun, 2004, 320(1):273-278
77.Hong SH,Gang EJ, Jeong JA, et al. In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells. Biochem Biophys Res Commun, 2005.330(4):1153-61
78. Philip N. Newsome U. Human cord blood-derived cells can differentiate into hepatocytes in the mouse . Liver with no evidence of celular fusion. Gastroenterology, 2003.124: 1891-1900.
79. Takahisa F, Tetsure H, Hideaki F, et al. Purification of adult hepatic progenitor cells using green fluorescent protein (GFP)-transgenic mice and fluorescence-activated cell sorting. J Hepatol, 2003, 39: 162-170
80. Mihonitou N, Yoshinori S, Katsutoshi I. Purification of Fetal Mouse Hepatoblasts by Magnetic Beads Coated with Monoclonal Anti-E-Cadherin Antibodies and Their In Vitro Culture. Exp Cell Res, 2002, 279: 330-343
81. Hisaya A,Tetsure H,Hideaki F,et al.Enrichment of Hepatic Progenitor Cells From Adult Mouse Liver. Hepatology, 2003. 37: 1385-1394
82. Atherine A, Luzaro EJ, Croager CM.et al.Establishment,characterization, and long-term maintenance of cultures of human fetal hepatocytes. Hepatology, 2003,38(5): 1095-1106
83. WeberA .Immortalization of hepatic Progenitor cells.Pathol Biol,2004;52(2):93-96
84. Dabeva MD,Petkov PM,Sandhu J, et al. Proliferation and differentiation of fetal liver epithelial progenitor cells after transplantation into adult rat liver. ANJ Pathol 2000:156:2017-2031
85. GermainL ,Noel M,Gourdeau H,et al.Promotion of growth and differentiation rat ductural oval cells in primary culture.Cancer Research.l988;46:368-378
86. Lazaro CA,Rhim JA,Yamada Y ,et al.Generation of hepatocytes from oval precursors in culture . Cancer Reasearch,1998;58:5514 - 5522.
87. Yang L,Li S,Hatch H,et al.Invitrotrans-differentiation of adult hepatic stem cells into pancreatic endocrine hormone -producing cells.Proc Natl Acad Sci USA .2002,99(12):8078 - 8083.
88. Suzuki A,Zheng YwYW,Kaneko S,et al.Clonal identification and characterization of self-renewing pluripotent stem cells in development liver.J Cell Biol.2002; 156(1): 173
89. Okano J,Shiota G,Matsumoto K.Hepatocyte growth factor exerts a proliferative effect on oval cells through the PI3K / AKT signaling pathway. Bioehem Biophys Res Commun,2003,309(2): 298-304
90. Matsusaka S, Tsujimura T, Toyosaka A, et al. Role of c-kit receptor tymsine kinase in development of oval cells in the rat 2-acetylaminofluorene / partial hepatectomy model.Hepatology. 1999,29: 670-676.
91. Knight B, Yeoh GCT, Husk KL'et al. Impaired preneoplatic changes and liver tumour form ation in tumour necrosis factor receptor type I knockout mice.J ExpMed, 2000, 192: 1809 1818
92. Overturf K; Al dhalimy M; OU CN; et al. Serial transplantation reveals the stem cell like regenerative potential of adult mouse hepatocytes. Am J Pathol, 1997, 151(5): 1273-1280.
93. Matsumoto K. Yoshitomih, Rossant J, et al. Liver organ ogenesis promoted by endothelial cells prior to vascular function. Science. 2001; 294(5542): 559-563.
94. Jung J, Zheng M, Goldfarb M, et al. Initiation of mammalian fiver development from endoderm by fibroblast growth factors. Science. 1999 ; 284(5422 :1998-2003.
95. Sund NJ,Ang SL,Sackett SD et al. Hepatocyte Nuclear Factor 3p (Foxa2) Is Dispensable for Maintaining the Differentiated State of the Adult Hepatocyte.Mol Cell Biol. 2000 July; 20(14): 5175-5183.
96. Zaret KS. Hepatocyte differentiation: from the endoderm and beyond. Curr Opin Genet Dev, 2001; 11(5): 568-574.
97. Eferl R, Sibilia M, Hilberg F, et al. Functions of c-Jun in fiver and heart development. J CeII Bio. 1999. 145 (51): 1049-1061.
98. Ying Zhang; Xue-Fan Bai; Chang-Xing Huang. Hepatic stem cells: existence and origin. World J Gastroenterol 2003; 9(2):201-204.
99. OmaryMB,Ku No,Toivoal DM, et al. Keratins: guardians of the liver. Hepatology; 2002; 35 (2): 251-256.
100. Heather AC,Stefan H,Luca F,et al. immunolocalization of putative human Liver progenitor cells in livers from patients with end-stage primary biliary cirrhosis and slerosing cholangitis using the monoclonal antibody OV-6.A m JP athol,1998,152:771-779.
101.Orla MC,Maura R,Gerry M, et al.In vitro evidence for the presence of hematopoietic stem cell in adult human liver.Hepatology,1999,29:1193-1198 .
102.Lucy GM,Michael PC,Nlamh N,et al.Differential express of lymphoid and myeloid markers on differenting hematopoietic stem cell in normal and tumor-bearing adult human liver.Hepatology,2000,31:1251-1256.
103.Ulrich B,Heather AC,Pamila R,et al.Expression of the stem cell factor receptor c-kit in normal and diseased pediatric liver:identification of a human hepatic progenitor cell?Hepatology, 1999,30:112-117.
104.Tan Y,Hughes D, Wang X, Costa RH. Adenovirus-mediated increase in HNF-3beta or HNF-3alpha shows differences in levels of liver glycogen and gene expression.Hepatology. 2002 Jan; 35(l):30-9.
105.Hughes DE; StolzDB; YuS; Tan Y; ReddyJK; WatkinsSC; Diehl AM; Costa RH. Elevated hepatocyte levels of the Forkhead box A2 (HNF-3beta) transcription factor cause postnatal steatosis and mitochondrial damage.Hepatology.2003 Jun; 37(6):1414-24.
106.Eleswarapu S,Jiang H.Growth hormone regulates the expression of hepatocyte nuclear factor-3 gamma and other liver-enriched transcription factors in the bovine liver.J Endocrinol. 2005 Jan; 184(l):95-105.
107.Westmacott A; Burke ZD; Oliver G; et al. C/EBPalpha and C/EBPbeta are markers of early liver development.Int J Dev Biol. 2006; 50(7):653-7.
108.Dontu G,A1 - Hajj M; Abdallah WM; et al.Stem cells in normal breast development and breast cancer. Cell Prolif; 2003:59- 72.
109.肖家诚;朱延波;朱上林;等。肝细胞肝癌中卵圆细胞的组织学与超微结构研究.临床与实验病理学杂志;2000;(16):177-179.
110.肖家诚,王天翔,Ruck P等。肝硬化组织中卵圆细胞的光镜、电镜与免疫电镜观察,J Clin Exp Pathol 2002;(1):57-60。
111.Shupe T; Petersen BE. Evidence regarding a stem cell origin of hepatocellular carcinoma.Stem Cell Rev. 2005; 1(3):261-4.
112. Alison MR. Liver stem cells: implications for hepatocarcinogenesis.Stem Cell Rev. 2005; 1(3):253-60.
113. Wu XZ,Yu XH. Bone marrow cells: the source of hepatocellular carcinoma? Med Hypotheses. 2007; 69(1):36-42.
114. Sell S, Pierce G B.Maturation arrest of stem cell differentiation is a common pathyway for the Cellular origin of teratocarcinomas and epithelial cancers.Lablnvest, 1994,70 (1):6-22
115. Hixson DC,Chapman L.McBride A,et al.Antigenic phenotypes common to rat oval cells,primary hepatocellular carcinoma sand developing bile ducts.Carcinogenesis, 1997,18 (6): 1169-1175
116.Sell S. Heterogeneity and plasticity of hepatocyte lineage cells.Hepatology,2001,33(3):738-750
117. Dumble M L,Croager E J, Yeoh G C T, et al.Generation and characterization of p53 null transformed hepatic progenitor cells:oval cells give rise to hepatocellular carcinoma.Carcinogenesis,2002,23(3):435-445
118. Bucher N L R, Farmer S R. Liver regeneration following partial hepatectomy: Genes And Metabolism.In: Stain A,Deihl AM(ed).Liver Growth And Repair.Chapman&Hall:London,l 998,1 -27
119. Thorgeirsson S S .Hepatic stem cells in liver regeneration.FASEBJ, 1996,10 (I1):1249-1256
120. Libbrecht L, Desmst Van Damme B, et al. Deep intralobular extension of human hepatic progenitor cells'correlates with parenchymal inflammation in chronic viral hepatitisxan progenit or cells'migrate?J Pathol,2000,15 6(3):1599-1612
121. Crosby H A, Hubscher S, Fabris L.et al. Immunolocalisation of putative human liver progenitor cells in liver of patients with end-stage primary biliary cirrhosis and sclerosing cholangitis using the antibody OV-6.A mJ P athol,l 998,152(3):771-779
122. Libbrecht L, Desmst V, Van Damme B, et al. Deep intralobular extention of human hepatic progenitor cells'crrelates with parenchymal inflammation in chronic viral hepatitisxan progenitor cells'migrate? J P athol,2000,192(3):373-378
123. Libbrecht L,De Vos R,Cassiman D,et al.Progenitor cells in hepatocellular adenomas.Am J Surg Pathol,2001,25(lt) :1388-1396
124. Lowes KN,Brennan B A ,Yeoh GC,et al.Oval cell numbers in human chronic disease are directly related to diseases severity.A m JP athol,19 99,15 4(2):537-541
125. Gong J Q, Fang C H. Relationship between the oval cells and development of hepatocellular carcinoma in rats.World Chin J Digestol,2002,10(10):1133-1139
126. Zulewski H,AbrahamE J,Gerlach M J,et al.Multipotential nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine,exocrine,and hepatic phenotypes.Diabetes,2001,50(3):521-533
127. WeissmanI L.Stem cells: units of development,units of regeneration,and units inevolution.Cell,2000,100(1):157-168
128 . Norris S,Collins C,Doherty D, et al.Resident heparie lymphocyres are phenotypically different from circulating lymphocytes. Hepatology,1998, 28:84-90.
129. Ruck-P,Xiao-J,Kaiserling-E.Small Epithlial cells and the histogenesis of hepatoblastoma.M ectron microscopic immunoelectron microscopic and immunohistochemical fingings.AmJ Pathol,1996,148:321-329.
130. Robrechts C,Devos R,Scoazec J,et al.Primary liver tumour of intermediate (hepatocyte-bileductcell)phenotype: a progenitor cell tumour?Liver, 1998,18:288 -293.
131. Kazushi S,Katsuya A,Takeshi I,et al .Expression of functional CD40 in hum anh epatocellularca rcinoma.Hepatology, 1999,30:920-926
132. A.M. James S; Camillo R; Bernhard J. International trial of the Edmonton protocol for islet transplantation.N Eng J Med; 2006; 355:1318-1330; 133. Perin EC; DohmannHF; Borojevic R; et al. Transendocardial; autologous bone marrow cell transplantation for severe; chronic ischemic heart failure . Circulation; 2003; 107 (18): 2294-2302.
134. Kumar KS; LefkowitchJ; RussoMW; et al. Successful sequential liver and stem cell transp lantation for hepatic due to p rimaryAlamyloidosis. Gastroenterology; 2003; 122 : 2026 - 2031.
135.姚鹏,胡大荣,王帅,等。自体骨髓干细胞移植治疗慢性重症肝病60例.实用医学杂志2005;19:2143-2145。
136.Lee SW,Wang X,Chowdhury NR, Hepatocyte transplantation: state of the art and strategies for overcoming existing hurdles. Ann Hepatol. 2004 Apr-Jun,3(2):48-53.
137.Ohashi K,Yokoyama T, Yamato M,et al. Engineering functional two- and three-dimensional liver systems in vivo using hepatic tissue sheets. Nat Med. 2007 Jul; 13(7):880-885.
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