摘要
目的分离和纯化人子宫内膜基质细胞和腺上皮细胞并进行体外培养,为建立胚胎体外种植模型奠定基础。方法通过酶消化、筛网过滤和差时贴壁等方法,分离和纯化人子宫内膜基质细胞和腺上皮细胞并继续体外培养,通过免疫细胞化学染色进行细胞表型鉴定。结果基质细胞和腺上皮细胞的产量分别为8×106个/g组织和6×106个/g组织,存活率分别为90%和85%,纯度分别达95%和90%;基质细胞波形蛋白染色阳性,腺上皮细胞角蛋白染色阳性;基质细胞接种后0.5 h开始贴壁,腺上皮细胞在接种24 h后贴壁;基质细胞可以有限的传代,腺上皮细胞不能传代。结论采用酶消化、筛网过滤和差时贴壁等方法,可获得纯度较高的子宫内膜基质细胞和腺上皮细胞。人子宫内膜细胞体外培养的成功为胚胎体外种植模型的建立奠定了基础。
Objective To set up an in-vitro model of embryo implantation by separating,purificating and culturing stromal and glandular epithelial cells of human endometriurn. Methods Human endometrial stromal and glandular epithelial cells were separated and purificated by enzymolysis, mesh filtration and pasted wall, then two types cells of purify were cultured Phenotype of stromal and glandular epithelial cells was identified by immunohistochemistry. Results The yields of stromal and glandular epithelia cells were 8 × 10^6/g and 6 × 10^6/g respectively, the surviving rate of two kinds cells were 90% and 85% respectively. Stromal cells were positive for vimenti, n, and glandular epithelial cells positive for cytokeratin. Stromal cells became adherent after 0. 5 h of culture, glandular cells became adherent after 24 h of culture. Stromal cell could be passaged for several generations, but glandular epithelial cells could not be passage＆ Conclusion highly purified strornal and glandular epithelial cells of human endometrium can be obtained by enzymolysis, mesh filtration and pasted wall purification. Successful culture in vitro of human endometrial cells lays a foundation for setting up an in-vitro model of embryo implantation.
出处
《贵州医药》
CAS
2007年第10期867-869,共3页
Guizhou Medical Journal
基金
贵州省重点攻关项目(计字2001-1214)
合肥市科委资助项目[合肥科合社字(2002)06号]
关键词
子宫内膜
基质细胞
腺上皮细胞
细胞培养
Human endometrium
Stromal cell
Glandular epithelial cell
Culture
