模拟子宫微环境诱导大鼠胎盘间充质干细胞向子宫内膜上皮细胞方向分化的实验研究

PMSCs to endometrial epithelial-like cells transformation under simulated uterine microenvironment in rats

摘要:

目的 模拟子宫微环境诱导大鼠胎盘间充质干细胞（PMSCs）向子宫内膜上皮细胞方向分化。方法 原代培养大鼠PMSCs及传代，绘制细胞生长曲线，制备子宫内膜条件培养液，用子宫内膜条件培养液和雌激素（β-雌二醇）进行体外诱导分化PMSCs；免疫荧光化学法检测细胞波形蛋白和角蛋白的表达；实时荧光定量聚合酶链反应（qRT-PCR）检测上皮细胞标记细胞角蛋白7（CK-7）、细胞角蛋白18（CK-18）、细胞角蛋白19（CK-19）、上皮膜抗原（EMA）的基因表达；Western blotting检测波形蛋白和角蛋白表达量。结果 实验组CK-7、CK-18、CK-19、EMA mRNA相对表达量高于对照组（P <0.05）；实验组波形蛋白相对表达量低于对照组（P <0.05）；实验组角蛋白相对表达量高于对照组（P <0.05）。结论 使用子宫内膜诱导培养液和雌激素能够促进大鼠PMSCs向子宫内膜上皮细胞方向分化，其作用显著，体外模拟子宫微环境在PMSCs向子宫内膜上皮细胞方向分化中有着重要的作用。

Abstract:

Objective To study the effect of simulated uterine microenvironment on the differentiation of rat placental mesenchymal stem cells (PMSCs) into endometrial epithelial cells.Methods Primary cultured rat PMSCs and passage, growth curve, endometrial conditioned medium, endometrial conditioned medium and estrogen (β-estradiol) to induce differentiation of PMSCs in vitro, immunofluorescence chemistry and Western blotting to detect the expression of vimentin and keratin in cells, qPCR to detect the gene table of epithelial cell markers cytokeratin 7 (CK-7), cytokeratin 18 (CK-18), cytokeratin 19 (CK-19), and epithelial membrane antigen (EMA).Results The relative expressions of CK-7, CK-18, CK-19, and EMRNA in experimental group were higher than those in control group (P < 0.05). The relative expression of vimentin in the experimental group was lower than that in the control group, and the relative expression of keratin in the experimental group was higher than that in the control group (P < 0.05).Conclusion The use of endometrial induction medium and estrogen can promote the differentiation of PMSCs into endometrial epithelial-like cells in rats, which plays an important role in the differentiation of PMSCs into endometrial epithelial-like cells in vitro.