基质胶对绒毛外细胞滋养层细胞体外侵袭的介导作用

Matrigel mediated role on extravillous cytotrophoblasts in vitro extracorporeal invasion

目的探讨基质胶（Matrigel）在绒毛外细胞滋养层细胞（EVCT）的体外侵袭中的作用。方法取健康早孕妇女（孕7～9周）人工流产绒毛组织,利用Percoll梯度离心法及差速贴壁法,将纯化的绒毛外细胞滋养层细胞,按所需浓度接种于涂有Matrigel培养板或24孔Transwell侵入系统中培养。用细胞免疫荧光法检测TGFβ2、cytokeration7、integrinα1β1。用CCK-8评价细胞生长状况。用Transwell细胞侵入系统检测EVCT的体外侵袭作用。结果①EVCT在Matrigel包被的Petri皿培养4h,用细胞免疫荧光法显示有95%以上的细胞表达TGFβ2、cytokeration7和integrinα1β1;②EVCT在Matrigel培养不同时间后,其生长指数未有明显变化,提示Matrigel对EVCT的生长未有明显影响;③EVCT在Transwell侵袭系统中培养不同时间后,侵袭细胞的OD值提示48h细胞侵袭作用达到平台期。结论 Matrigel在EVCT体外侵袭中并不影响其细胞纯度及其生长曲线,其介导的体外侵袭在48h达到侵入平台期。

Objective To investigate matrigel mediated effect on extracorporeal invasion of extravillous cytotrophoblasts （EVCT）. Methods Taking abortion villi of the healthy pregnant women （pregnancy 7-9 weeks）, using Percoll gradient centrifugation and differential adhesion method, according to the concentration required for immunization the purified extravillous trophoblast cells were vaccinated in the Matrigel-coated culture plates or 24hole culture Transwell invasion system. By using the cell-mediated immunity assay TGFβ2, cytokeration 7, integrin α1β1 were detected; using CCK-8 cell growth conditions was evaluated; using transwell cell systems with intrusion the in vitro invasionrole of EVCT was detected. Result 1） EVCT cultured in the matrigel-coated Petri Min for 4 h showed that more than 95% of the cells expressed TGFβ2, cytokeration 7, integrin α1β1. 2） EVCT at different times in the matrigel culture, its growth index had no significant change, suggesting Matrigel no significant effect on the growth. 3） EVCT cultured in Transwell invasion system at different time the OD values of invasive cells suggested that the cell invasive role at 48 h reached plateaustage. Conclusion Matrigel in EVCT extracorporeal invasion has not affecting these cell purity and their growth curve, the mediated extracorporeal invasion at 48 hours can reach platform stage.