早孕绒毛细胞滋养层细胞分离培养的实验研究  

In vitro culture of human cytotrophoblasts isolated from early pregnancy villi

作　　者：蒋立艳[1] 李真[1] 成娅[1] 陈波洁[1] 王智[1] 林桂兰[1] 刘萍[1] 杨合荣[2] 

机构地区：[1]第三军医大学新桥医院妇产科,重庆400037 [2]第三军医大学西南医院妇产科,重庆400038

出　　处：《第三军医大学学报》2004年第14期1248-1251,共4页Journal of Third Military Medical University

摘　　要：目的　改善体外分离和培养人早孕绒毛细胞滋养层细胞 (cytotrophoblast,CTB)的方法 ,探讨CTB的一些生物学特性。方法　取人工流产 6～ 8周妊娠绒毛 ,机械法分离 ,酶消化 ,经Ficoll分离介质离心得到单个核细胞进行培养并鉴定 ,测定其生长曲线并观察其体外转化、细胞周期和激素合成。结果　分离培养的CTB经免疫细胞化学染色鉴定 ,接种后2h开始贴壁 ,第 5天融合 90 %以上 ,细胞周期显示约有 79%细胞处于G0 G1 期 ,激素分泌表现出随CTB的转化而变化。结论　成功分离培养了人早孕绒毛CTB ,方法简便易行 ,获得的细胞形态单一、生长稳定、定向转化 。Objective To improve the methods for isolation and culture of human cytotrophoblasts (CTBs) from early pregnancy villi and to investigate its partial biological properties. Methods Human villi from 6-8 weeks legal voluntary interruption pregnancy were mechanically isolated, enzyme digested and dispersed into mononuclear cells by Ficoll gradient centrifugation. The mononuclear cells were cultured and identified, and their growth curve was determined. Other biological properties such as transition in vitro , cell cycle, and hormone synthesis were observed. Results Immunocytochemistry revealed that CTBs began to adhere to the wall at 2 h after inoculation, while more than 90% cells fused at 5 d. Cell cycle revealed that 79% cells were at G0/G1. The secretion of E2, β HCG, and PROG increased in a CTB dependent manner. Conclusion We have successfully cultured human cytotrophoblasts isolated from early pregnancy villi and obtained monomorphic cells with stable growth, which provides a useful experimental technique for the studies of the roles of CTBs in reproductive physiology.

关 键 词：细胞滋养层细胞 细胞培养 生物学特性 

分 类 号：R321.4[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]