人子宫内膜细胞原代体外培养方法

Establishment of Method for Human Endometrial Cell Culture in Vitro

目的改良人子宫内膜细胞原代体外培养方法的建立,并对其进行特征鉴定。方法将选择的子宫内膜组织机械法分离,复合酶消化,传代自然增殖法纯化细胞,进行单层培养。通过倒置显微镜观察其大体形态,HE染色观察其胞核情况,免疫细胞化学染色法对间质细胞(endometrial stromal cell,ESC)及上皮细胞(endometrial glandular epithelial cell,EEC)进行鉴定。结果培养的子宫内膜细胞均有ESC和EEC两种形态细胞。EEC和ESC经角蛋白单抗体和波形蛋白单抗体染色,凡是胞质被染成棕黄色的细胞为阳性细胞。结论成功培养了原代人子宫内膜细胞,方法经济、简单,获得细胞成活率高,生长稳定。成功建立子宫内膜细胞原代体外培养体系,为从细胞和分子水平方面研究子宫内膜的各种功能及干预调节奠定基础。

Objective To improve the method for endometrial cell culture in vitro and identification of the cells. Methods Human endometrial tissues were separated mechanically, digested by complex enzyme, purified by passage and monolayer cultured. The cellular morphology was observed by inverted phase contrast microscope, the nucleus and cytoplasm were observed by staining of haematoxylin and eosin （HE）; Endometrial stromal cell and endometrial glandular epithelial cell were identified by the method of immunocytochemistry of cytokeratin （CK18） and vimentin （Vim）. Rusults Both endometrial stromal cell （ESC） and endometrial glandular epithelial cell （EEC） were existed in the culture; EEC and ESC stained by CK18 and Vim McAb in the brown-yellow were positive cells. Conclusion The endometrial cell is cultured successfully in vitro and the method is established with high survival rate and low cost, which contributes to investigate endometrial function and adjust in cell and molecular level.