两种人早孕绒毛滋养层细胞的原代培养方法

目的:比较两种不同方法进行人早孕绒毛滋养层细胞原代培养的异同。方法:分别用组织块培养法和3种酶联合消化法(Ⅳ型胶原酶、低浓度胰酶及DNaseⅠ)、Ficoll 400梯度密度离心法对人早孕绒毛滋养层细胞培养。结果:两种方法进行早孕滋养层细胞的培养均获得成功,其生长率比较差异无显著性意义(P>0.05)。结论:组织块培养法简单易行,但细胞数量少,生长缓慢,难以完成后续实验;3酶联合消化、Ficoll 400梯度密度离心法相比较而言,可获得数量更多的活力好、纯度较高的具有不同分化功能的滋养层细胞,为进一步研究其侵袭、增殖能力等生理、病理作用提供了坚实的基础。

Study on two primary culture of human first trimester cytotrophoblasts

Objective:To explore the difference of two isolation and primary culture of human first trimester cytotrophoblasts.Methods:We isolated and cultured human first trimester cytotrophoblasts from early pregnancy villous using two different methods in vitro.One method was tissue cultivation,the other method followed as:cytotrophoblasts was mechanically isolated,digested by three enzymes,and purified by Ficoll 400 gradient centrifugation.Results:We succeed to isolate human first trimester cytotrophoblasts from early pregnancy villous by two primary culture methods.Conclusion:Comparing to tissue cultivation,the method which cytotrophoblast is mechanically isolated,digested by three enzymes,and purified by Ficoll 400 gradient centrifugation can be obtained more cytotrophoblasts with powerful activity and high purity in vitro,these cytotrophoblasts have different differentiated functions and provided us a useful experimental technique for further study of their physiological and pathological roles such as their invasiveness,proliferate activity.