早孕期人细胞滋养层细胞的分化及分离、培养、鉴定  

Isolation and Differentiation of Human First Trimester Trophoblasts

作　　者：王海燕[1] 归绥琪[1] 邹琴娣[1] 李雪莲[1] 

机构地区：[1]复旦大学附属妇产科医院,上海200011

出　　处：《生殖与避孕》2008年第4期215-221,共7页Reproduction and Contraception

基　　金：上海市科委上海市现代生物与药业基金(No.02D219115);复旦大学985工程资助项目

摘　　要：目的:分离正常早孕期人细胞滋养层细胞并分析其分化功能。方法:用不同的消化方法分离正常早孕期人细胞滋养层细胞,分别在体外培养24h,用相差显微镜、Giemsa染色、免疫细胞化学法、扫描电镜、Transwell分析其形态及分化功能。结果:①采用低浓度胰酶一次性消化法获得绒毛外滋养层细胞,特异表达细胞角蛋白7,不表达波形蛋白,细胞互相聚集但不聚合,具有高度增殖活性和侵袭能力;②采用高浓度胰酶、长时间连续消化法分离得到绒毛滋养层细胞,具有高度融合、分泌hCG-β的合体滋养层细胞特性;③采用低浓度胰酶连续消化法获得的细胞包含绒毛外滋养层细胞和绒毛滋养层细胞,其共性是表达细胞总角蛋白,不表达波形蛋白。结论:利用不同的分离方法可有效、经济地获得具有不同分化功能的早孕期人细胞滋养层细胞。Objective： To isolate human first trimester cytotrophoblasts and discuss their differentiation in vitro. Methods： Human first trimester cytotrophoblasts were isolated using different digestive conditions. After 24 h culture in vitro, cells morphology and function were observed by discrepancy microscopy, Giemsa dye, immunocytochemistry, transwell invasive assay and scanning electron microscopy. Results： （1） The cell populations expressed cytokeratin 7 but not vimentin obtained by low and once enzymatic digestion without vibration. These cells aggregated but never fused in Matrigel or rat-tail gel. Cells had the proliferative and invasive properties of extravillous cytotrophoblasts. （2） The remaining villi were digested by sequential high enzymatic digestion. These cells had the characteration of villous cytotrophoblasts （VCT）. VCT fused to form mutiplenucleated syncytiotrophoblasts which expressed hCG-β but not vimentin. （3） Cells obtained by low and sequential enzymatic digestion were implantated in the plate covered by Matrigel or rat-tail gel. These cells contained two types of cytotrophoblasts which expressed cytokeratin but not vimentin. Conclusion： Different isolation methods might obtain different cytotrophoblasts with different characteristics and functions.

关 键 词：绒毛外细胞滋养层细胞(EVCT) 绒毛细胞滋养层细胞(VCT) 侵袭 增殖 融合 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]