摘要
目的:建立人早孕期绒毛细胞滋养细胞(villous cytotrophoblasts,VCT)及绒毛外细胞滋养细胞(extravillous cytotrophoblasts,EVCT)的分离、培养方法。 方法:利用不同的胰酶消化条件,收集人早孕期绒毛组织的VCT及EVCT并分别培养。倒置显微镜、扫描电镜观察VCT及EVCT的形态学特征;免疫细胞化学鉴定细胞来源及纯度。 结果:胰酶短期、温和消化获得的EVCT,可生长于Matrigel包被的培养器皿上,并表达特异性标志物c-crbB-2;延长消化时间、增加胰酶浓度获得的VCT,种植于塑料或玻璃培养器皿上可聚集并融合形成合体滋养细胞。VCT不表达c-crbB-2。结论:采用不同的胰酶消化及体外培养条件,可简单、快捷地分别获得高纯度人早孕期VCT及EVCT。
Objective: To establish a method to isolate and purify villous cytotrophoblasts (VCT) and extravillous cytotrophoblasts (EVCT) from the chorionic villi of the first trimester human placenta. Methods: When cytotrophoblast cells were selectively released under different tissue disaggregation conditions, two types of trophoblast cells were collected. Immunocytochemistry and electron microscopy revealed the various characteristics of these cells. Results: EVCT could be isolated with gentle enzymatic digestion while VCT isolation required more intense disaggregation conditions. EVCT cultured on Matrigel exhibited positive staining for cytokeratin and c erbB-2, whereas VCT cultured on plastic dishes aggregated and fused to form syncytiotrophoblasts and showed no immunoreative c-erbB-2. Conclusion: This method can be successfully used to isolate pure EVCT and VCT.
出处
《生殖与避孕》
CAS
CSCD
北大核心
2004年第2期70-73,81,T001,T002,共7页
Reproduction and Contraception
基金
复旦大学"985工程"项目资助
