C-erbB_2蛋白在小鼠围着床期子宫中的表达及其对体外胚胎着床影响的研究
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摘要
目的:探索C-erbB_2蛋白在小鼠围着床期子宫组织中的表达与定位;研究小鼠子宫内膜上皮(endometrial epithelial cells EECs)细胞的分离、鉴定和体外培养技术;建立小鼠体外胚胎着床的模型和共培养体系;初步探讨c-erbB_2反义寡核苷酸(c-erbB_2 ASODN)对小鼠体外胚胎着床的影响。
方法:1、用促超排法制作妊娠小鼠的动物模型;2、用免疫组化法观察C-erbB_2蛋白在围着床期小鼠子宫组织中的表达与定位;3、用分步酶消化法分离培养EECs,并用抗细胞角蛋白(CK-19)免疫细胞化学方法鉴定EECs;4、将已贴壁的EECs与获取的小鼠胚泡进行共培养以建立小鼠体外胚胎着床模型,分组观察比较胚泡在不同培养体系中的脱带率、粘附率、扩展率,确定理想的共培养体系;5、分组观察比较c-erbB_2 ASODN对小鼠体外胚胎着床的影响;6、用western blotting法检测各组EECs中C-erbB_2蛋白的表达情况。
结果:1、免疫组化结果显示在未孕子宫和空白对照组子宫中未见C-erbB_2蛋白的表达,D1-3天子宫中C-erbB_2蛋白主要分布在子宫内膜腔上皮和腺上皮细胞胞膜上,子宫基质细胞中未见表达,D4-5天子宫中C-erbB_2蛋白除了表达在子宫内膜腔上皮和腺上皮细胞胞膜上,上皮下基质细胞膜上可见弱阳性表达,D6-8天的子宫中C-erbB_2蛋白主要分布在子宫内膜腔上皮和腺上皮细胞以及围绕着床位点的蜕膜细胞胞膜上,各组肌层均未见阳性表达;2、分步酶消化法成功分离培养EECs,用抗CK-19免疫细胞化学染色鉴定EECs,其阳性率达到90%以上;3、三组共培养组(无血清共培养、3%FBS共培养组、0.4%BSA共培养组)的脱带率、粘附率和扩展率明显高于胚泡单独培养组(P<0.05),3%FBS共培养组和0.4%BSA共培养组的脱带率、粘附率和扩展率又明显高于无血清共培养组(P<0.05),而3%FBS共培养组的脱带率、粘附率、扩展率和0.4%BSA共培养组相比,差别无显著性(P>0.05);4、用c-erbB_2 ASODN干预着床模型结果显示:各组24小时脱带率差别无统计学意义(P>0.05);5μmol/L c-erbB_2 ASODN组48小时粘附率与扩展率低于对照组,但差别无统计学意义(P>0.05),10μmol/L c-erbB_2 ASODN组48小时粘附率与扩展率明显低于对照组,差别有统计学意义(P<0.05),15、20μmol/L c-erbB_2 ASODN组48小时粘附率与扩展率显著低于对照组,差别有显著统计学意义(P<0.01),而20μmol/L c-erbB_2 ODN组48小时的粘附率和扩展率与对照组相比无差别(P>0.05); 5、Western Blotting法检测c-erbB_2 ASODN各组EECs中C-erbB_2蛋白表达结果显示:不同浓度的c-erbB_2 ASODN作用着床模型48小时后,5μmol/L c-erbB_2 ASODN组蛋白的表达低于对照组,但差别无统计学意义(P>0.05), 10μmol/L c-erbB_2 ASODN组表达明显低于对照组,差别有统计学意义(P<0.05),15、20μmol/L c-erbB_2 ASODN组蛋白的表达显著低于对照组,差别有显著统计学意义(P<0.01);另外,20μmol/L c-erbB_2 ASODN作用24小时后C-erbB_2蛋白表达低于对照组(P<0.05),20μmol/L c-erbB_2 ASODN作用48小时后C-erbB_2蛋白表达显著低于对照组(P<0.01)。
结论:1、C-erbB_2蛋白在着床期小鼠子宫组织中表达有规律性变化;2、分步酶消化法是分离培养子宫内膜上皮细胞的有效方法;3、胚泡与EECs共培养是比较理想的着床模型,3%FBS和0.4%BSA都是比较理想的共培养体系;4、C-erbB_2蛋白能促进小鼠胚胎的体外着床。
Objective:To investigate the expression and distribution of C-erbB_2 protein in the periimplantation mouse uterus; to research the separation, cultivation and identification technology of endometrial epithelial cells(EECs) in vitro; to build the mouse implantation model in vitro and find the ideal cultivational system of mouse implantation model; To preliminarily investigate the effection of c-erbB_2 ASODN on the mouse implantation model in vitro.
Methods:1、The pro-ovulated method was used to make pregnant animal models; 2、Immunohistochemistry staining was used to examine the localization and expression of the C-erbB_2 protein in the peri-implantation uterus; 3、The EECs was divided from the Day 4 pregnant uterus by enzyme digestion, and the percentage of the EECs expressing CK-19 was more than 90%; 4、We used man-made glass pipes to inhale the mice blastocysts, and cultivated them with the EECs, in order to determine the ideal co-cultivated system then we divided them into 4 groups; 5、We used the contradict method to observe the effects of the c-erbB_2 ASODN on mouse implantation model in vitro, and divided them into 6 groups,after 24 hours’co-culture we recorded the hatching rate of these groups, after 48 hours’co-culture we recorded the adhesion rate and outgrowth rate of these groups; 6、The expression of C-erbB_2 albumen in the EECs was detected by Western Blotting.
Results:1、Immunohistochemistry experiments showed that unique uterine cell-specific C-erbB_2 protein distribution. The nagetive control slice and non-pregnant uterus exhibited no signals. On day 1-3 unlike in the stromal cells, the C-erbB_2 protein was detected primarily in the EECs; on day 4-5 besides the EECs, the stromal cells expressed a little C-erbB_2 protein, on day 6-8 the EECs and the decidualized cells around the implanting blastocyst exhibited the accumulation of C-erbB_2 protein. 2、The percentage of the EECs expressing CK-19 was 90% more or less , which were isolated from the mouse day 4 pregnant uterus. 3、As shown in the blank the ratios of embryonic hatchment, attachment and outgrowth of the BSA group and the FBS group were obviously higher than the before two groups (P<0.05), but the difference between the last two groups were no significant (P>0.05);There was no difference in the ratios of embryonic hatchment in six groups; 4、The ratios of embryonic attachment and outgrowth between the c-erbB_2 ODN group and control group were no difference either(P>0.05), the ratio of embryonic attachment and outgrowth in group including 5μmol/L c-erbB_2 ASODN is a little lower than control group(P>0.05), the ratios of 48 hours’embryonic attachment and outgrown in groups including 10、15、20μmol/L c-erbB_2 ASODN were obviously lower than control group (P<0.05、P<0.01); 5、The expression of C-erbB_2 protein in EECs was detected by Western blotting. the level of C-erbB_2 protein in groups including 5 c-erbB_2 ASODN was lower than control group, but the difference has no significance( P>0.05), and the level of C-erbB_2 protein in groups including 10 c-erbB_2 ASODN was lower than control group, the differnence has significance(P<0.05), the level of C-erbB_2 protein in groups including 15、20μmol/L c-erbB_2 ASODN was lower than control group, the difference has big significance (P<0.01); the level of C-erbB_2 protein in groups including 20 c-erbB_2 ASODN acting for 24 hours and 48 hours was lower than control group(P<0.05).
Conclusions:1、The expression of C-erbB_2 protein in the periimplantation mouse uterus changes regularly; 2、The method of enzyme digestion is a valid method to separate the EECs from the Day 4 pregnant mouse uterus; 3、The implantation models of blastocysts cultivated with EECs in medium including FBS and BSA are both the ideal implantation models; 4、C-erbB_2 protein could promote mouse embryo implantation in vitro.
方法:1、用促超排法制作妊娠小鼠的动物模型;2、用免疫组化法观察C-erbB_2蛋白在围着床期小鼠子宫组织中的表达与定位;3、用分步酶消化法分离培养EECs,并用抗细胞角蛋白(CK-19)免疫细胞化学方法鉴定EECs;4、将已贴壁的EECs与获取的小鼠胚泡进行共培养以建立小鼠体外胚胎着床模型,分组观察比较胚泡在不同培养体系中的脱带率、粘附率、扩展率,确定理想的共培养体系;5、分组观察比较c-erbB_2 ASODN对小鼠体外胚胎着床的影响;6、用western blotting法检测各组EECs中C-erbB_2蛋白的表达情况。
结果:1、免疫组化结果显示在未孕子宫和空白对照组子宫中未见C-erbB_2蛋白的表达,D1-3天子宫中C-erbB_2蛋白主要分布在子宫内膜腔上皮和腺上皮细胞胞膜上,子宫基质细胞中未见表达,D4-5天子宫中C-erbB_2蛋白除了表达在子宫内膜腔上皮和腺上皮细胞胞膜上,上皮下基质细胞膜上可见弱阳性表达,D6-8天的子宫中C-erbB_2蛋白主要分布在子宫内膜腔上皮和腺上皮细胞以及围绕着床位点的蜕膜细胞胞膜上,各组肌层均未见阳性表达;2、分步酶消化法成功分离培养EECs,用抗CK-19免疫细胞化学染色鉴定EECs,其阳性率达到90%以上;3、三组共培养组(无血清共培养、3%FBS共培养组、0.4%BSA共培养组)的脱带率、粘附率和扩展率明显高于胚泡单独培养组(P<0.05),3%FBS共培养组和0.4%BSA共培养组的脱带率、粘附率和扩展率又明显高于无血清共培养组(P<0.05),而3%FBS共培养组的脱带率、粘附率、扩展率和0.4%BSA共培养组相比,差别无显著性(P>0.05);4、用c-erbB_2 ASODN干预着床模型结果显示:各组24小时脱带率差别无统计学意义(P>0.05);5μmol/L c-erbB_2 ASODN组48小时粘附率与扩展率低于对照组,但差别无统计学意义(P>0.05),10μmol/L c-erbB_2 ASODN组48小时粘附率与扩展率明显低于对照组,差别有统计学意义(P<0.05),15、20μmol/L c-erbB_2 ASODN组48小时粘附率与扩展率显著低于对照组,差别有显著统计学意义(P<0.01),而20μmol/L c-erbB_2 ODN组48小时的粘附率和扩展率与对照组相比无差别(P>0.05); 5、Western Blotting法检测c-erbB_2 ASODN各组EECs中C-erbB_2蛋白表达结果显示:不同浓度的c-erbB_2 ASODN作用着床模型48小时后,5μmol/L c-erbB_2 ASODN组蛋白的表达低于对照组,但差别无统计学意义(P>0.05), 10μmol/L c-erbB_2 ASODN组表达明显低于对照组,差别有统计学意义(P<0.05),15、20μmol/L c-erbB_2 ASODN组蛋白的表达显著低于对照组,差别有显著统计学意义(P<0.01);另外,20μmol/L c-erbB_2 ASODN作用24小时后C-erbB_2蛋白表达低于对照组(P<0.05),20μmol/L c-erbB_2 ASODN作用48小时后C-erbB_2蛋白表达显著低于对照组(P<0.01)。
结论:1、C-erbB_2蛋白在着床期小鼠子宫组织中表达有规律性变化;2、分步酶消化法是分离培养子宫内膜上皮细胞的有效方法;3、胚泡与EECs共培养是比较理想的着床模型,3%FBS和0.4%BSA都是比较理想的共培养体系;4、C-erbB_2蛋白能促进小鼠胚胎的体外着床。
Objective:To investigate the expression and distribution of C-erbB_2 protein in the periimplantation mouse uterus; to research the separation, cultivation and identification technology of endometrial epithelial cells(EECs) in vitro; to build the mouse implantation model in vitro and find the ideal cultivational system of mouse implantation model; To preliminarily investigate the effection of c-erbB_2 ASODN on the mouse implantation model in vitro.
Methods:1、The pro-ovulated method was used to make pregnant animal models; 2、Immunohistochemistry staining was used to examine the localization and expression of the C-erbB_2 protein in the peri-implantation uterus; 3、The EECs was divided from the Day 4 pregnant uterus by enzyme digestion, and the percentage of the EECs expressing CK-19 was more than 90%; 4、We used man-made glass pipes to inhale the mice blastocysts, and cultivated them with the EECs, in order to determine the ideal co-cultivated system then we divided them into 4 groups; 5、We used the contradict method to observe the effects of the c-erbB_2 ASODN on mouse implantation model in vitro, and divided them into 6 groups,after 24 hours’co-culture we recorded the hatching rate of these groups, after 48 hours’co-culture we recorded the adhesion rate and outgrowth rate of these groups; 6、The expression of C-erbB_2 albumen in the EECs was detected by Western Blotting.
Results:1、Immunohistochemistry experiments showed that unique uterine cell-specific C-erbB_2 protein distribution. The nagetive control slice and non-pregnant uterus exhibited no signals. On day 1-3 unlike in the stromal cells, the C-erbB_2 protein was detected primarily in the EECs; on day 4-5 besides the EECs, the stromal cells expressed a little C-erbB_2 protein, on day 6-8 the EECs and the decidualized cells around the implanting blastocyst exhibited the accumulation of C-erbB_2 protein. 2、The percentage of the EECs expressing CK-19 was 90% more or less , which were isolated from the mouse day 4 pregnant uterus. 3、As shown in the blank the ratios of embryonic hatchment, attachment and outgrowth of the BSA group and the FBS group were obviously higher than the before two groups (P<0.05), but the difference between the last two groups were no significant (P>0.05);There was no difference in the ratios of embryonic hatchment in six groups; 4、The ratios of embryonic attachment and outgrowth between the c-erbB_2 ODN group and control group were no difference either(P>0.05), the ratio of embryonic attachment and outgrowth in group including 5μmol/L c-erbB_2 ASODN is a little lower than control group(P>0.05), the ratios of 48 hours’embryonic attachment and outgrown in groups including 10、15、20μmol/L c-erbB_2 ASODN were obviously lower than control group (P<0.05、P<0.01); 5、The expression of C-erbB_2 protein in EECs was detected by Western blotting. the level of C-erbB_2 protein in groups including 5 c-erbB_2 ASODN was lower than control group, but the difference has no significance( P>0.05), and the level of C-erbB_2 protein in groups including 10 c-erbB_2 ASODN was lower than control group, the differnence has significance(P<0.05), the level of C-erbB_2 protein in groups including 15、20μmol/L c-erbB_2 ASODN was lower than control group, the difference has big significance (P<0.01); the level of C-erbB_2 protein in groups including 20 c-erbB_2 ASODN acting for 24 hours and 48 hours was lower than control group(P<0.05).
Conclusions:1、The expression of C-erbB_2 protein in the periimplantation mouse uterus changes regularly; 2、The method of enzyme digestion is a valid method to separate the EECs from the Day 4 pregnant mouse uterus; 3、The implantation models of blastocysts cultivated with EECs in medium including FBS and BSA are both the ideal implantation models; 4、C-erbB_2 protein could promote mouse embryo implantation in vitro.
引文
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[39] Jing Li, Jian V. Zhang, Yu-Jing Cao, et al. Inhibition of the β-catenin Signaling Pathway in Blastocyst and Uterus during the Window of Implantation in Mice[J]. Biology of Reproduction, 2005,72(3): 700-706
[40] Suzana Atanasoski, Steven S.Scherer, Erich Sirkowski, et al. ErbB2 Signaling in Schwann Cells Is Mostly Dispensable for Maintenance of Myelinated Peripheral Nerves and Proliferation of Adult Schwann Cells after Injury[J]. The Journal of Neuroscience, 2006, 26(7): 2124-2131
[41] Hideho Ueda, Akihiro Oikawa, Akinori Nakamura, et al. Neurgulin Receptor ErbB2 Localization at T-tubule in Cardiac and Skeletal Muscle[J]. Journal of Histochemistry and Cytochemistry, 2005, 53(1): 87-91
[42] Sang-Oh Yoon, Sejeong Shin, Elizabeth A.Lipscomb. A Novel Mechanism for Integrin-Mediated Ras Activation in Breast Carcinoma Cells: The α6β4 Integrin Regulates ErbB2 Translation and Transactives Epidermal Growth Factor Receptor/ErbB2 Signaling[J]. Cancer Res, 2006, 66(5):2732-2739
[43] Christian Spangenberg, Ekkehart U.Lausch, Tatjana M.Trost, et al. ErbB2-Mediated Transcripitional Up-regulation of the α5β1 Integrin Fibronectin Receptor Promotes TumorCell Survival Under Adverse Conditions[J]. Cancer Res, 2006;66(7): 3715-3725
[44] Victoria P.Ramsauer, Vanessa Pino, Amjad Farooq, et al. Muc4-ErbB2 Complex Formation and Signaling Polarized CACO-2 Epithelail Cells Indicate That Muc4 Acts as an Unorthodox Ligand for ErbB2[J]. Molecular Biology of the Cell, 2006, 17:2931-2941
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[47] Paria BC, Tsukamura H, Dey SK. Epidermal Growth Factor-specific Protein Tyrosine Phosphorylation in Preimplantation Embryo Development[J]. Biol Reprod, 1991, 45(5):711-8
[48] Cao JP, Yu JK, Li C, et al. Integrin beta-1 is Involved in the Signaling of Glial Cell Line-derived Neurotrophic Factor[J]. J Comp Neurol, 2008, 509(2):203-210
[49] Junwen Qin, Laura D-C, Juan-Enrique Schwarze, et al. Effects of Progranulin on Blastocyst Hatching and Subsequent Adhesion and Outgrowth in the Mouse[J]. Biol Reprod, 2005, 73: 434-442
[50] 王绍娟, 谌兵来, 顾勉琴等. 整合素 β3 反义寡核苷酸抗小鼠妊娠的作用. 复旦学报, 2005, 32(1): 75-78
[51] 李南, 魏莎莉,杨戎等. sLex 在小鼠胚泡植入早期子宫内膜中的表达及作用. 生殖与避孕, 2007, 27(8): 497-50
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[39] Jing Li, Jian V. Zhang, Yu-Jing Cao, et al. Inhibition of the β-catenin Signaling Pathway in Blastocyst and Uterus during the Window of Implantation in Mice[J]. Biology of Reproduction, 2005,72(3): 700-706
[40] Suzana Atanasoski, Steven S.Scherer, Erich Sirkowski, et al. ErbB2 Signaling in Schwann Cells Is Mostly Dispensable for Maintenance of Myelinated Peripheral Nerves and Proliferation of Adult Schwann Cells after Injury[J]. The Journal of Neuroscience, 2006, 26(7): 2124-2131
[41] Hideho Ueda, Akihiro Oikawa, Akinori Nakamura, et al. Neurgulin Receptor ErbB2 Localization at T-tubule in Cardiac and Skeletal Muscle[J]. Journal of Histochemistry and Cytochemistry, 2005, 53(1): 87-91
[42] Sang-Oh Yoon, Sejeong Shin, Elizabeth A.Lipscomb. A Novel Mechanism for Integrin-Mediated Ras Activation in Breast Carcinoma Cells: The α6β4 Integrin Regulates ErbB2 Translation and Transactives Epidermal Growth Factor Receptor/ErbB2 Signaling[J]. Cancer Res, 2006, 66(5):2732-2739
[43] Christian Spangenberg, Ekkehart U.Lausch, Tatjana M.Trost, et al. ErbB2-Mediated Transcripitional Up-regulation of the α5β1 Integrin Fibronectin Receptor Promotes TumorCell Survival Under Adverse Conditions[J]. Cancer Res, 2006;66(7): 3715-3725
[44] Victoria P.Ramsauer, Vanessa Pino, Amjad Farooq, et al. Muc4-ErbB2 Complex Formation and Signaling Polarized CACO-2 Epithelail Cells Indicate That Muc4 Acts as an Unorthodox Ligand for ErbB2[J]. Molecular Biology of the Cell, 2006, 17:2931-2941
[45] Florina Haimovici and Deborah J. Anderson. Effects of Growth Factors and Growth Factor-Extracellular Matrix Interactions on mouse Trophoblast Outgrowth in vitro[J]. Biology of Reproduction, 1993,49(1): 124-130
[46] Fouladi-Nashta AA, Jones CJ, Nijjar N, et al. Characterization of the Uterine Pheotype during the Peri-implantation Period for LIF-null, MF1 Strain Mice[J]. Dev Biol, 2005, 281(1): 1-21
[47] Paria BC, Tsukamura H, Dey SK. Epidermal Growth Factor-specific Protein Tyrosine Phosphorylation in Preimplantation Embryo Development[J]. Biol Reprod, 1991, 45(5):711-8
[48] Cao JP, Yu JK, Li C, et al. Integrin beta-1 is Involved in the Signaling of Glial Cell Line-derived Neurotrophic Factor[J]. J Comp Neurol, 2008, 509(2):203-210
[49] Junwen Qin, Laura D-C, Juan-Enrique Schwarze, et al. Effects of Progranulin on Blastocyst Hatching and Subsequent Adhesion and Outgrowth in the Mouse[J]. Biol Reprod, 2005, 73: 434-442
[50] 王绍娟, 谌兵来, 顾勉琴等. 整合素 β3 反义寡核苷酸抗小鼠妊娠的作用. 复旦学报, 2005, 32(1): 75-78
[51] 李南, 魏莎莉,杨戎等. sLex 在小鼠胚泡植入早期子宫内膜中的表达及作用. 生殖与避孕, 2007, 27(8): 497-50
[1] 史小林.人类生殖学[M].北京:科学出版社,2002.41.
[2] Vigano P, Blasio AMD, Deu’Antonio G, e1 a1. Culture of Human Endometrial Cells: A New Simple Technique to Completely Separate Epithelial Glands[J]. Acta Obstet Gynecol Scand, 1993 Feb, 72(2):87—92 .
[3] Overton C E, Barlow D, Shaw S F, et al, Hicks B: In vitro Culture of Endometrial Stromal and Gland Cells as A Model for Endometriosis: the Effect of Peritoneal Fluid on Proliferation[J]. Fertil Steril 1997 Jan, 67(1):51-56.
[4] Seli E, Arici A. Endometriosis: in Teraction of Immune and Endocrine Systems[J]. Sem in Reprod Med, 2003, 21(2):135-144.
[5] Hombach-Klonisch S, Kehlen A, fowler P A, et al. Regulation of Functional Steroid Receptors and Ligand-Induced Responses in Telomerase-Immortalized Human Endomertrial EpithelialCells[J]. J Mol Endocrinol, 2005, 34(2):517-534.
[6] Dimitriadis E, Stoikos C, Baca M, et al. Relaxin and Prostaglandin E2 Regulate Interleukin 11 during Human Endometrial Stromal Cell Decidualization.[J]. J Clin Endocrinol Metab, 2005, 90(6):3458-3465.
[7] Grant KS, Wira CR. Effect of Mouse Uterine Stromal Cells on Epithelial Cell Transepithelial Resistance (TER) and TNF-α and TGF-β Release in Culture[J]. Biology of Reproduction, 2003 May, 69:1091-1098.
[8] Krikun G, Mor G, Lockwood C, The Immortalization of Human Endometrial Cells[J]. Methods Mol Med. 2006, 121:79-83.
[9] Wei P,Tao SX, Zhang XS, et al. Effect of RU486 on Apoptosis and P53 Expression at the Boumdary of Fetal-Maternal Interface of Rhesus Monkey (Macaca Mulatta)[J]. Sheng Li Xue Bao, 2004, 56(1):60-65.
[10] Wei P, Yuan J X, Jin X, et al. Molsidomine and N-omega-nitro-L-arginine Methyk Ester Inhibit Implanatation and Apposes in Mouse Endometrium[J]. ActaPharmaco sin, 2003, 24(12):1177-1184.
[11] 朱槿,周剑萍,贺斌等.C-myc基因mRNA在小鼠胚胎着床期间的表达[J].《生殖与避孕》,2001,21(2):77-81.
[12] Kyo S, Nakamura M, Kiyono T, et al. Successful Immortalization of Endometrial Glandular Cells with Normal Structural and Functional Characteristics[J]. Am J Pathol, 2003, 163(6): 2259-2269.
[13] Gu Y, Gibori G, Rat Decidual Cell Cultures[J]. Methods Mol Med, 2006, 121:69-78.
[14] Ryan I P, Schriock E D , Taylor RN. Isolation, Characterization, and Comparison of Human Endometrial and Endometriosis Cells in Vitro[J]. J Clin Endocrinol Metab. 1994 Mar, 78(3):642—649.
[15] Selam B, Arici A. Implantation defect in endometriosis: endometrium or peritoneal fluid[J]. J Reprod Fertil Suppl, 2000; 55:121-128.
[16] Matthews C J , Redfern C P , Hirst B H , et a1. Characterization of Human Purified Epithelial and Stromal Cells from Endometrium and Endometriosis in Tissue Culture[J]. Fertil Steril, 1992 May, 57( 5):990-997.
[17] Sharpe-Timms K L. Defining Endometrial Cells:the Need for Improved Identification at Ectopic Sites and Characterization in Eutopic Sites for Developing Novel Methods of Management for Endometriosis[J]. Fertil Steril, 2005 Jul, 84(1):35-37.
[18] Dimitriadis E, Robb L, Salamonsen L A. Interleukin 11 Advances Progestrone-Induced Decidualization of Human Endometrial Stromal Cells[J]. Mol Hum Reprod, 2002 Jul, 8(7):636-643.
[19] Dimitriadis E, Stoikos C, Baca M, et al. Relaxin and Prostaglandin E2 Regulate Interleukin 11 During Human Endometrial Stromal Cell Decidualization [J]. J Clin Endocrinol Metab, 2005, 90(6):3458-3465.
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