人早孕期蜕膜基质细胞及蜕膜腺上皮细胞的分离培养方法

Isolation,Culture and Identification of Decidual Stromal Cells and Decidual Epithelial Cells of Human First-Trimester Pregnancy

目的:建立一种经济、高效的人早孕期蜕膜基质细胞及蜕膜腺上皮细胞分离、培养方法。方法:胶原酶、胰酶联合消化蜕膜组织,网筛过滤、密度梯度离心及差速贴壁法纯化细胞;免疫细胞化学法鉴定细胞纯度;MTT增殖实验分析细胞体外增殖活力。结果:利用此法可成功体外培养蜕膜基质细胞及蜕膜腺上皮细胞,细胞纯度可达90%以上,并在体外呈现良好的生长状态。结论:此法经济、高效,能够同时获得纯度较高的蜕膜基质细胞及蜕膜腺上皮细胞。

Objective： To establish a method for isolation,purification and culture of decidual stromal cells and decidual epithelial cells of human first-trimester pregnancy.Methods： Decidual tissues were digested with 0.1% Collagenase Ⅳ and 0.25% trypsin.Mesh screen filtration,density gradient centrifugation and differential adhesion methods were used to isolate and purify the cells.Two types of cells were identified by immunocytochemistry with vimentin and cytokeratin-7.MTT proliferation assay was employed to analyze the viability of these two kinds of cells in vitro.Results： Decidual stromal cells and decidual epithelial cells could be isolated and cultured successfully in vitro with this method.The cell purity was over 90% and the viability of cultured cells was further confirmed by MTT proliferative assay.Conclusion： Decidual stromal cells and decidual epithelial cells could be colleted at the same time with this economical and efficient method.