太行青山羊附睾头细胞系的建立
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摘要
试验旨在获得太行青山羊附睾头细胞体外培养体系,为附睾特异表达蛋白功能研究提供材料。对15日龄太行青山羊进行无菌去势获得附睾头组织,剪碎组织后,通过胰蛋白酶和胶原酶I消化水解,经过原代培养、传代培养、细胞冻存和复苏对细胞状态进行初步鉴定,另外绘制了生长曲线并分析了染色体数目。结果表明:原代培养、传代培养、冻存复苏后细胞状态良好,2~3 d能够达到90%融合度,且细胞生长趋势整体呈"S"型,染色体数目为2n=60,数目正常。该研究成功建立了太行青山羊附睾头细胞系。
The aim of this research was to establish epithelial cells of the epididymal caput in Taihang black goat in order to provide materials for the study of specific-epididymal protein.The epididymis caput was obtained from 15-day-old goats steriled and castrated. The tissue was digested and hydrolyzed by trypsin and collagenase I. The epithelial cell status was preliminary identified after primary culture,subculture,cell cryopreservation and resuscitation. The growth curve and chromosome number analysis were also carried out. The results showed that the epithelial cells were normally with 90% confluency in 2 to 3 days in primary culture,subculture and cryopreservation,and the cell growth trend was S curve in general. The number of chromosomes was 2 n=60,the number was normal. This study successfully established epithelial cell lines of caput in Taihang black goat.
The aim of this research was to establish epithelial cells of the epididymal caput in Taihang black goat in order to provide materials for the study of specific-epididymal protein.The epididymis caput was obtained from 15-day-old goats steriled and castrated. The tissue was digested and hydrolyzed by trypsin and collagenase I. The epithelial cell status was preliminary identified after primary culture,subculture,cell cryopreservation and resuscitation. The growth curve and chromosome number analysis were also carried out. The results showed that the epithelial cells were normally with 90% confluency in 2 to 3 days in primary culture,subculture and cryopreservation,and the cell growth trend was S curve in general. The number of chromosomes was 2 n=60,the number was normal. This study successfully established epithelial cell lines of caput in Taihang black goat.
引文
[1]胡向农,张忠林,杨建军.大鼠附睾上皮细胞的体外培养与研究[J].南京铁道医学院学报,1999(1):3-5.
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[4]Breton S,Ruan Y C,Park Y J,et al.Regulation of epithelial function,differentiation,and remodeling in the epididymis[J].Asian J Androl,2016,18(1):3-9.
[5]郭丽娜,张国林,任有蛇,等.β防御素104a在不同日龄雄性山羊睾丸与附睾中的定位及表达特性[J].山西农业大学学报(自然科学版),2017(12):866-871.
[6]马红.大白猪仔猪皮肤成纤维细胞系建立及生物学特征[J].华北农学报,2009(S1):96-98.
[7]范晓梅,张通,栗瑞兰,等.绒山羊附睾上皮细胞培养方法的建立及其生长特性[J].畜牧兽医学报,2017(7):1212-1220.
[8]杨森,冯付明,王银辉.乳兔成骨细胞原代培养与鉴定:改良胶原酶与胰酶的分段消化[J].中国组织工程研究,2014(38):6129-6135.
[9]陈东风,刘为纹,房殿春,等.人胆囊上皮细胞的分离、培养及鉴定[J].第三军医大学学报,2001(9):1109-1111.
[2]石慧,李建远.附睾上皮细胞体外培养模式综述[J].中外医学研究,2011(9):120-121.
[3]刘海涛,舒端阳.动物细胞培养问题研究[J].实验教学与仪器,2017(Z1):86-87.
[4]Breton S,Ruan Y C,Park Y J,et al.Regulation of epithelial function,differentiation,and remodeling in the epididymis[J].Asian J Androl,2016,18(1):3-9.
[5]郭丽娜,张国林,任有蛇,等.β防御素104a在不同日龄雄性山羊睾丸与附睾中的定位及表达特性[J].山西农业大学学报(自然科学版),2017(12):866-871.
[6]马红.大白猪仔猪皮肤成纤维细胞系建立及生物学特征[J].华北农学报,2009(S1):96-98.
[7]范晓梅,张通,栗瑞兰,等.绒山羊附睾上皮细胞培养方法的建立及其生长特性[J].畜牧兽医学报,2017(7):1212-1220.
[8]杨森,冯付明,王银辉.乳兔成骨细胞原代培养与鉴定:改良胶原酶与胰酶的分段消化[J].中国组织工程研究,2014(38):6129-6135.
[9]陈东风,刘为纹,房殿春,等.人胆囊上皮细胞的分离、培养及鉴定[J].第三军医大学学报,2001(9):1109-1111.