人胚胎成纤维细胞的培养及饲养层的制备  

Culture of human embryonic fibroblasts and production of feeder layers for human spermatogonial stem cells

作　　者：王毓斌[1] 陈斌[1] 张志玲[1] 王颖超[1] 王鸿祥[1] 向祖琼[1] 卢永宁[1] 胡凯[1] 杨怡珂[2] 韩银发[1] 汪铮[3] 王益鑫[1] 黄翼然[1] 

机构地区：[1]上海交通大学医学院附属仁济医院泌尿外科上海市男科学研究所,上海200001 [2]上海交通大学医学院附属仁济医院妇产科 [3]上海市消化疾病研究所干细胞实验室

出　　处：《中国男科学杂志》2008年第10期21-24,共4页Chinese Journal of Andrology

摘　　要：目的从人胚胎躯干中分离、培养人胚胎成纤维细胞,建立人胚胎成纤维细胞饲养层用于人精原干细胞的培养。方法利用酶消化法从孕5～9周龄人胚胎躯干中分离培养人胚胎成纤维细胞,制作饲养层,使用不同浓度丝裂霉素C处理,以细胞形态、生长曲线作为胚胎成纤维细胞和饲养层的评价指标。结果从人胚胎中成功分离培养出人胚胎成纤维细胞,该细胞可传代15代以上,且经过传代及冷冻复苏后生物学特性无改变。12.5mg/L丝裂霉素C作用2h可达到较好处理效果。结论成功分离和培养人胚胎来源的成纤维细胞,用于人精原干细胞饲养层的制作。Objective To isolate and cultivate human embryonic fibroblasts（hEFs） derived from human embryos, trunks and to establish feeder layer culture system of it for culturing of human spermatogonial stem cells. Methods The hEFs were isolated by enzyme digestion from the subcutaneous connective tissue of 5- to 9- week old human embryos. Different concentrations of mitomycin C were used to pretreat feeder layer, The effects of hEFs and feeder layers were assessed by cell morphology and growth curves. Results The hEFs were successfully isolated and cultivated from human embryos, trunks. They could be passaged beyond the 15th generation. The biologic characteristics of the cells did not change, even in high-passage cells or frozen-thawed cells.The optimal concentration of mytomycin C to inhibit proliferation of hEFs was 12.5mg/L for 2h. Conclusion The hEFs derived from human embryos, trunks are successfully isolated and cultivated, and to product feeder layers for human spermatogonial stem cells.

关 键 词：成纤维细胞 胚胎 细胞培养技术 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]