人正常子宫内膜原代腺上皮细胞的优化分离和培养

目的 优化人正常子宫内膜腺上皮细胞分离和原代培养的方法,提供子宫内膜相关疾病研究的体外细胞模型。方法 采用酶消化、筛网过滤、离心的方法体外分离和培养人子宫内膜腺上皮与间质细胞。腺上皮细胞鉴定采用光学显微镜下观察细胞形态；免疫细胞荧光及免疫细胞化学法检测上皮细胞角蛋白(CK)和间质细胞波形蛋白(Vim)的表达。结果 经诊刮获得的18份标本中有17份分离培养成功；细胞形态符合腺上皮细胞特征,CK免疫荧光及免疫化学染色阳性,Vim染色阴性,纯度达90%以上；正常原代人子宫内膜腺上皮细胞不能传代,培养5～6d后细胞逐渐衰老。结论 改良后的原代培养方法取材简单,克服了污染问题,可获得高纯度及足够数量的人正常子宫内膜腺上皮细胞作为体外实验模型。
Objective To develop an optimized method for separation and primary culture of normal human endometrial epithelial cells. Methods The human endometrial epithelial cells were isolated with enzymatic digestion, after filtration and centrifugation the cells were identified by light microscopy and immunocytochemistry/immunofluorescence staining with cytokeratin and vimentin monoclonal antibodies.Results The primary culture of human endometrial epithelial cells was successfully establi-shed in 17 out of 18 endometrial samples from biopsy specimens. The morphology of the cells maintained the biologic characteristics of endometrial glandular epithelial cells.The cells were stained positively with cytokeratin and negatively with vimentin (purity＞90%). The glandular epithelial cells could not be passaged, and the cells aging after 5-6d. Conclusion The modified method developed in this study can be used to obtain high purity and adequate normal endometrial glandular epithelial cells for study of endometrial diseases