人子宫在位和异位内膜间质细胞的分离纯化和体外培养

Isolation,purification and cultivation in vitro of stromal cells in eutopic endometrium and ectopic endometrium of human uterus

目的:建立人子宫正常内膜、子宫内膜异位症(内异症)在位及异位内膜间质细胞的分离、培养的方法。方法:将12例正常子宫内膜、9例在位子宫内膜及6例异位子宫内膜经分段酶解、筛网过滤、沉降及贴壁纯化技术进行分离、纯化和体外培养、光镜观察,应用鼠抗人波形蛋白抗体及鼠抗人细胞角蛋白单克隆抗体对间质细胞进行免疫细胞化学染色鉴定。结果:10例正常子宫内膜、6例在位子宫内膜及4例异位子宫内膜标本分离、培养成功,间质细胞纯度均可达93%以上,并均可传6～8代。结论:采用分段酶解、筛网过滤、沉降及贴壁纯化的方法可获得纯度较高的子宫内膜间质细胞,为进一步研究内异症发生、发展的分子生物学机制提供了理想的实验模型。

Objective：To establish the method of isolation and cultivation of stromal cells in eutopic endometrium and ectopic endometrium of cases with endometriosis and normal endometrium of healthy cases.Methods：Isolation, purification and cultivation in vitro were performed by segment enzymatic hydrolysis, screening filtration, sedimentation and adherent purification technique;observation under light microscope, mouse anti-human vimentin antibody, mouse anti-human cytokeratin monoclonal antibody were used for immunocytochemical staining and identification among 12 samples of normal endometrium, 9 samples of eutopic endometrium and 6 samples of ectopic endometrium.Results：Isolation and cultivation of 10 samples of normal endometrium, 6 samples of eutopic endometrium and 4 samples of ectopic endometrium were successful, the purity of stromal cells was up to 93%, and all the stromal cells could be transferred for six to eight generations.Conclusion：Segment enzymatic hydrolysis, screening filtration, sedimentation and adherent purification technique can obtain uterine stromal cells with high purity, which provide ideal experimental model for further investigating the molecular biological mechanism of occurrence and development of endometriosis.