人子宫内膜基质细胞的分离纯化和体外培养方法研究  

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

作　　者：高红艳[1,2] 陈继明[2] 何援利[1] 

机构地区：[1]南方医科大学附属珠江医院妇产科,广州510280 [2]苏州大学附属第三医院妇产科,江苏常州213003

出　　处：《重庆医学》2014年第34期4634-4636,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(81270658)

摘　　要：目的分离和纯化人子宫内膜基质细胞用于宫腔粘连发病机制的研究。方法通过二次酶消化、一次网筛和差时贴壁等方法,分离和纯化人子宫内膜基质细胞,体外培养传代,通过免疫组织化学法进行细胞表型鉴定。结果基质细胞在接种后30min开始贴壁,可见梭形和多角形2种形态的细胞。免疫组织化学显示这2种细胞具有波形蛋白免疫反应性,反应率可达95%以上,而细胞角蛋白无免疫反应性,提示其为来源于内膜基质的基质细胞。分离出的子宫内膜基质细胞可以在体外进行增殖。结论采用二次酶消化、一次网筛和差时贴壁纯化等方法,可成功分离人子宫内膜基质细胞,且基质细胞可以有限的传代。Objective To isolate,purify human endometrial stromal cells,and culture these cells in vitro,for the further research of pathogenesis of intrauterine adhesions.Methods Human endometrial stromal cells were separated and purificated by enzymo ly-sis,mesh filtration and pasted wall,then the stromal cells of purification were cultured.Phenotype of stromal cells was identified by immunohistochemistry.Results The stromal cells became adherent 30 min after culture.Two types of stromal cells were found. One was spindle and the other was polygonal.Immunocytochemical staining method revealed that vimentin was positive in both types of stromal cells,and the positive rate was above 9 5%;meanwhile,cytokeratin was negative in both types of stromal cells, which indicated that these cells were endometrial stromal cells.The isolated endometrial stromal cells can be cultured in vitro.Con-clusion Highly purified stromal cells of human endometrium could be obtained by enzymolysis,mesh filtration and pasted wall pur-ification,and the cultured stromal cells could passage to limited generations.

关 键 词：子宫内膜基质细胞 细胞培养技术 荧光免疫测定 宫腔粘连 

分 类 号：R711[医药卫生—妇产科学]