小鼠子宫内膜上皮细胞的分离和原代培养

Isolation and primary culture of uterus endometrial cells of mice

目的建立一种高效的子宫内膜上皮细胞分离和体外培养方法,为子宫疾病的发病机制或治疗药物筛选的进一步研究提供一个比较理想和有价值的的实验模型。方法用酶消化、过滤、离心与差速贴壁纯化相结合的方法分离培养小鼠子宫内膜上皮细胞,以上皮细胞角蛋白为抗原的免疫荧光法对分离培养的细胞进行纯度鉴定。结果小鼠子宫内膜上皮细胞培养4～5 h后细胞可以贴壁生长;24 h时细胞即可生成单层,排列紧密,呈长、圆或多角形生长,呈角蛋白阳性,胞质呈棕色,核呈蓝色,纯度达95%以上。结论该方法能够成功分离并得到高产量、高纯度、高增殖能力的子宫内膜上皮细胞。

Objective To establish an efficient method of separation and culture of mouse uterus endometrial epithelial cell in vitro,and to provide an ideal and valuable experimental model for further study of the pathogenesis of uterine disease or drug screening.Methods Uterus endometrial epithelial cells were isolated and purified by digestion of trypsin,centrifugation and differential adherence,and observed by light microscope and identified by immunocytochemical stain,and the purity was also analyzed by immumofluorescence method with cytokeratin as the antigen.Results High purity epithelial cells（95%） were obtained;the epithelial cells became adherent after 4-5 hours ＇culture,formed monolayer cell colony after culturing 24 hours and were positively stained by anti-cytokeratin antibody;the cells presented egg-shape,the cytoplasm showed red color,and the nuclus blue.Conclusion Mouse uterus endometrial cells of high outcome,high purity and high proliferation can be obtained by this method.