雌激素与孕激素在兔子宫内膜上皮细胞体外增殖中的作用  

Effect of β-estradiol and progesterone on the proliferation of endometrial epithelial cells in vitro

作　　者：段翠密[1] 王海滨[1] 江红[1] 王常勇[1] 宋宇轩[1] 王和平[1] 郭希民[1] 李晶[1] 项婷婷[1] 

机构地区：[1]军事医学科学院基础医学研究所组织工程研究室,北京100850

出　　处：《解放军医学杂志》2006年第6期582-584,共3页Medical Journal of Chinese People's Liberation Army

基　　金：全军"十五"医药卫生科研基金课题(06MA318)

摘　　要：目的探讨雌激素E2和孕激素P4不同水平对家兔子宫内膜上皮细胞体外增殖的影响。方法离心法获取家兔子宫内膜上皮细胞;用上皮角蛋白抗体的免疫荧光细胞染色对细胞进行鉴定,用流式细胞仪检测上皮细胞的纯度。设不同雌激素和孕激素水平组合添加于培养液,利用MTT法间接测定上皮细胞的体外增殖能力。结果利用离心法可获得高纯度的子宫内膜上皮细胞(96%);雌激素(100nmol/L)和少量孕激素(10nmol/L)对上皮细胞增殖有促进作用。免疫荧光细胞检测结果表明,经激素刺激的子宫内膜上皮细胞对上皮角蛋白抗体反应呈阳性。结论性腺激素对体外培养的子宫内膜上皮细胞有促进增殖的作用,而且不会使其分化为其他类型细胞。Objective The purpose of this study was to investigate the effect of different level of sex steroids β-estradiol and progesterone） on the proliferation of uterus endometrial epithelial cells. Methods Rabbit uterus endometrial epithelial cells were isolated by digestion of trypsin and centrifugation. The specificity and homogeneity of the endometrial epithelial cells cultured in media containing β-estradiol and progesterone were determined by immunofluorescence staining of cytokeratin monoclonal antibody and the flow cytometric analysis. The different concentration of progesterone （P4） and β-estradiol （E2） were added in media, the proliferative capability of the endometrial epithelial cells cells in vitro was determined indirectly by MTT assay. Results High purity endometrial epithelial cells （96%） were isolated using centrifugation method. The endometrial epithelial cells were examined by immunofluorescence staining. The endometrial epi- thelial cells were positive stained by anti-cytokeratin antibody. Immunofluorescence staining demonstrated that cytokeratin was detectable in the cytoplasm of epithelial cells. The cell growth curve showed that the proliferation of endometrial epithelial cells was not significantly changed by the treatment of E2 （100nmol/L） or P4 （100nmol/L） individually, but a combination of 100nmol/L E2 and 10nmol/L P4 could promote its proliferation greatly （P〈0. 05）. We culture the endometrial epithelial cells in medium contains 100nmol/L E2 and 10nmol/L P4 and the endometrial epithelial cells can be sub-cultured 7-8 generations and they even can be cultured after recovery from cryopreservation. Conclusion Our results indicated that sex steroids （E2 and P4） could promote proliferation of endometrial epithelial cells in vitro and maintain its specificity and homogeneity.

关 键 词：子宫内膜 雌激素 孕激素 细胞增殖 

分 类 号：R318[医药卫生—生物医学工程]