摘要
目的建立一种高效分离子宫内膜上皮细胞与间质细胞的方法。方法收集全子宫切除患者的健康子宫内膜组织,将子宫内膜组织剪碎后,采用二次酶消化-二次过滤-差时贴壁的方法将人子宫内膜间质细胞(HUSC)与上皮细胞(HUEC)分离。采用流式细胞仪鉴定分离所得人子宫内膜基质和上皮细胞;并进一步对原代子宫内膜间质细胞进行诱导蜕膜化。结果流式细胞术检测分离的间质细胞波形蛋白阳性率为(97.0±2.5)%,上皮细胞角蛋白阳性率为(90.0±4.1)%。此外,使用环磷酸腺苷(c AMP)联合醋酸甲羟孕酮(MPA)体外诱导子宫内膜间质细胞产生了良好的蜕膜化反应。结论成功建立了一种高效分离原代人子宫内膜基质和上皮细胞的方法。
Objective To establish an efficient method for isolating primary human endometrium stromal and epithelial cells in vitro.Methods Health endometrial tissue of hysterectomy patient was obtained,and was cut into a small piece.Firstly,the human endometrial stromal cells (HUSC) and human endometrial epithelial cells (HUEC) were isolated by the twice enzymatic digestion,twice separation and different adherent time of the two types of cells.And then HUSC and HUEC were identified and the percentage of vimentin (Vim)-positive and cytokeratin (CK)-positive ones were determined by flow cytometry.After that,primary HUSC were stimulated by cyclic adenosine monophosphate (cAMP)+medroxyprogesterone acetate (MPA) for 4 d and 7 d.Results The HUSC purity was (97.0±2.5)% and the HUEC purity was (90.0 ± 4.1)% determined by flow cytometry,indicating that we had highly purified cultures of HUSC and HUEC in vitro.After cAMP+MPA induction for 4 d and 7 d,the HUSC showed more obvious changes in cell morphology and prolactin (PRL) level.Conclusion We successfully established an efficient method of isolation and characterization of primary human endometrium stromal and epithelial cells in vitro.
出处
《中华生殖与避孕杂志》
CAS
CSCD
北大核心
2017年第2期124-128,共5页
Chinese Journal of Reproduction and Contraception
基金
科技基础性工作专项(2013FY110521)
国家自然科学青年基金(81601343)
扬州市自然基金(YZ2016110)~~
关键词
人子宫内膜间质细胞
人子宫内膜上皮细胞
分离方法
标志分子
蜕膜化
Human endometrium stromal cells
Human endometrium epithelial cells
Isolation method
Molecular markers
Decidualization