人早孕蜕膜组织子宫内膜间质干细胞向子宫内膜上皮细胞的分化  

Endometrial stromal stem cells from early-pregnancy decidua differentiate into endometrial epithelial cells

作　　者：陈波[1] 

机构地区：[1]菏泽市立医院产科,山东省菏泽市274031

出　　处：《中国组织工程研究》2016年第14期2098-2103,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:蜕膜组织是重要的子宫内膜干细胞来源之一,从蜕膜组织中可分离出间充质干细胞。通过对其进行一定的诱导,可分化为不同的细胞类型。目的:从人早孕蜕膜组织中分离得到子宫内膜间质干细胞,对其进行诱导分化,观察其向子宫内膜上皮细胞分化的情况。方法:获得人早孕蜕膜组织,进行子宫内膜间质干细胞分离培养和鉴定。对子宫内膜间质干细胞进行定(向分化诱,进行实验分组,分别为实验组子宫内膜条件培养液)和对照组(含胎牛血清的正常培养基),利用免疫荧光法检测角蛋白表达情况。并进行Western bolt检测,检测实验组与对照组细胞的角蛋白相对表达量。结果与结论:(1)从人早孕蜕膜组织中成功分离培养获得子宫内膜间质干细胞,细胞体外生长曲线呈"S"形,接种后1-48 h,曲线平缓上升,细胞处于适应期。接种48 h之后,曲线陡峭上升,细胞增殖速度加快,进入对数期;接种60 h之后,细胞生长速度下降,曲线平稳上升;(2)经流式细胞仪检测,子宫内膜间质干细胞可以表达CD73、CD90,不表达CD34、CD45、细胞角蛋白;(3)经免疫荧光检测,角蛋白表达方面,对照组不表达,实验组大部分细胞均呈阳性表达;(4)经Western bolt检测,实验组细胞的角蛋白相对表达量显著高于对照组(P<0.05);(5)实验结果表明,从人早孕蜕膜组织中可以分离获得子宫内膜间质干细胞,通过对细胞进行定向诱导,可以使其向子宫内膜上皮细胞发生分化。BACKGROUND： Decidua tissue is one of the most important sources of endometrial stem cells, from which stromal stem cells can differentiate into various kinds of cells under contain inductions. OBJECTIVE： To observe the differentiation of endometrial stromal stem cells isolated from early-pregnancy decidua into endometrial epithelial cells.METHODS： Endometrial stromal stem cells from early-pregnancy decidua were cultured, identified and directionally induced. Then induced cells were randomized into test group cultured in conditioned medium and control group cultured in normal culture medium containing fetal bovine serum. Expression of keratin was detected by immunofluorescence assay and western bolt assay in the two groups. RESULTS AND CONCLUSION： The endometrial stromal stem cells from early-pregnancy decidua were successfully isolated and cultured, and presented with a S-shaped curve. Within 1-48 hours after inoculation, cultured cells were in the adaptation phase; then, the cells proliferated rapidly and began to grow logarithmically at 48 hours after culture; after 60 hours of culture, the cell growth rate decreased, with a steady rise in the cell curve. Results from the flow cytometry detection showed that the endometrial stromal stem cells could express CD73 and CD90, but not express CD34, CD45 and cytokeratin. Immunofluorescence findings showed the expression of keratin negative in the control group, and mostly positive in the test group. Besides, the relative expression of keratin in the test group was significantly higher than that in the control group（P〈 0.05）. These results suggest that the endometrial stromal stem cells can be isolated from the human decidua in early pregnancy, and differentiate into endometrial epithelial cells by directional induction.

关 键 词：子宫 干细胞 蜕膜 组织工程 培养 子宫内膜干细胞 子宫内膜 间质干细胞 蜕膜组织 细胞分化 上皮细胞 

分 类 号：R394.2[医药卫生—医学遗传学]