小鼠子宫内膜基质细胞的分离纯化与体外培养

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

目的分离和纯化小鼠子宫内膜基质细胞并进行体外培养,为进一步研究子宫内膜异位症的发病机制提供实验基础。方法通过酶消化、滤网过滤和差时贴壁等方法 ,分离和纯化小鼠子宫内膜基质细胞,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0.5 h开始贴壁,可见梭形和多角形两种形态的细胞。免疫组化显示这两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,提示它们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。结论采用酶消化、滤网过滤和差时贴壁等方法 ,可成功地分离小鼠子宫内膜基质细胞,基质细胞可以有限的传代。

Objective To isolate, purify mouse endometrial stromal cells , and culture these cells in vitro , which might lay a foundation for the further research of pathogenesis of endometriosis .Methods Mouse endometrial stromal cells were separated and purificated by enzymo lysis , 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 after 0.5h of culture.Two shapes of stromal cells were found .One was spindle and the other was polygonal .Immunocytochemical staining method showed that both shapes of stromal cells were positive for vimentin , with above 95%positive rate, and negative for cytokeratin , indicating that these cells were endometrial stromal cells .The isolated endometrial stromal cells can be cultured in vitro .Conclusion Highly purified stromal and cells of mouse endometrium can be obtained by enzymolysis , mesh filtration and pasted wall purification .Stromal cell could be limitedly passaged for several generations .Successful culture in vitro of mouse endometrial stromal cells lays a foundation for the further study of endometriosis .