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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 ,为进一步研究CTB在生殖生理中的作用提供了实验基础
Objective To improve the isolation and culture of human gestational trophoblastic cells (CTB) in vitro in vitro and to explore some biological characteristics of CTB. Methods Human chorionic villi were collected from 6 to 8 weeks after artificial abortion. The cells were isolated by mechanical method and digested by enzyme. The mononuclear cells were cultured and identified by centrifugation in Ficoll medium. The growth curve was measured and the in vitro transformation, cell cycle and hormone synthesis were observed. Results CTBs were identified by immunocytochemistry. The cells adhered to the surface 2h after inoculation and over 90% on the 5th day. The cell cycle showed that about 79% of cells were in the G0G1 phase, and the hormone secretion showed changes with the transformation of CTB . Conclusion CTB of human chorionic villi was successfully isolated and cultured in this study. The method is simple and easy to perform. The obtained single cell morphology, stable growth and directional transformation provide the experimental basis for further study on the role of CTB in reproductive physiology.