人子宫内膜基质细胞及腺上皮细胞的分离纯化和体外培养

Isolation, purification and culture in vitro of glandular epithelial cell and stromal cells of human endometrium

目的:建立人子宫正常内膜、内膜异位症在位及异位子宫内膜的基质及腺上皮细胞分离、培养的方法,为进一步研究子宫内膜异位症发生、发展的分子生物学机制提供一个理想的实验模型。方法:15例正常子宫内膜、15例在位子宫内膜及10例异位内膜经胶原酶消化、筛网过滤、差时贴壁等技术进行分离、纯化和体外培养,光镜观察,应用鼠抗人波形蛋白抗体、鼠抗人细胞角蛋白单克隆抗体免疫组化染色对间质及上皮细胞进行鉴定。结果:正常子宫内膜和子宫内膜异位症在位子宫内膜标本分离、培养均成功;5例异位内膜标本获得成功,基质细胞和腺上皮细胞纯度均可达95%以上,并均可传代。结论:采用酶解、筛网分离及贴壁能获得纯度较高的基质与腺上皮细胞。人子宫内膜细胞分离体外培养的成功,有助于研究子宫内膜异位症的发病机制,为临床实验提供重要的理论依据。

Objective:To establish a method of isolation, purification and culturing of human glandular epithelial and stromal cells of normal, eutopic, and ectopic endometrium in vitro. Methods:Enzymolysis, filtration, and pasted wall purification were used to isolate and purify the glandular epithelial and stromal cells in 15 normal endometrium, 15 eutopic endometrium and 10 ectopic endometrium. Two types of cells were observed by light microscopy and verified by immunocytochemical staining with vimentin and cytokeratin. Results:Endometrial cell culture was successfully established in all normal endometrium and eutopic endometrium of enmetriosis, but in 5 of 10 ectopic endometrium of endometriosi. The purities of stromal cells and glandular epithelial cells were 95% and 92% respectively, glandular epithelial cell and stromal cells all can passage for several generations. Conclusion:Highly purified epithelial and stromal cells can be obtained through enzymolysis, filtration, and pasted wall purification. The cellular model will be useful to investigate the pathogenesis of endometriosis.