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目的探究原代人子宫在位内膜细胞分离及培养的简易方法,比较分离培养单一间质细胞和混合培养间质细胞之间的差异。方法采用单纯胰蛋白酶,分段消化人子宫内膜组织,经过两次筛网过滤、离心纯化技术,分离、纯化人子宫内膜间质细胞(ESC)和腺上皮细胞(EEC),采用免疫荧光方法对细胞进行鉴定,采用Cell Titer96~ Aqueous One Solution Reagent(MTS)方法监测细胞之间增长速度。结果间质细胞多呈梭形或多角形,纯度可达97%以上。腺上皮细胞多似蝌蚪形或多角形,纯度可达93%以上。混合培养细胞比单一培养细胞增殖能力强。结论采用单纯胰蛋白酶分段消化法和二次筛网过滤法可成功分离子宫内膜腺上皮细胞和间质细胞,具有获得细胞成活率高、生长稳定的优点,混合培养间质细胞,保留细胞间整体性,可为进一步研究子宫内膜细胞提供一定的实验基础。
Objective To explore a simple method for isolation and culture of eutopic endometrial cells in primary human uterus, and to compare the differences between single stromal cells and mesenchymal cells isolated and cultured. Methods Human pancreatic endometrial tissue was digested with trypsin and purified twice by centrifugation and sieving twice. Human endometrial stromal cells (ESCs) and glandular epithelial cells (EEC) were isolated and purified by immunofluorescence Methods The cells were identified and the cell growth rate was monitored by Cell Titer96 ~ Aqueous One Solution Reagent (MTS). Results Interstitial cells mostly fusiform or polygonal, purity up to 97%. Glandular epithelial cells like tadpoles or polygons, purity up to 93%. Mixed culture cells than single cultured cells proliferation ability. Conclusions The endometrial glandular epithelial cells and stromal cells can be successfully separated by simple trypsin digestion and secondary sieve filtration. It has the advantages of high cell survival rate and stable growth. Mixed culture of mesenchymal cells and retained cells Between the integrity, for further study of endometrial cells provide a certain experimental basis.