体外诱导脂肪来源干细胞向黏膜上皮细胞分化的研究.PDF

　 军 杂 2015年10月1 日　第40卷　第10期 798 解放 医学 志　 论　著 体外诱导脂肪来源干细胞向黏膜上皮细胞分化的研究 王程，徐潇，陈冲，李梁，闫俊灵，汤苏阳 [摘要]　目的　探讨在体外诱导脂肪来源干细胞(ADSCs)分化为黏膜上皮细胞的可能性。方法　获取成年人皮下 脂肪组织，进行ADSCs 的分离、培养，扩增至第三代进行细胞免疫表型、分化情况及遗传稳定性鉴定。分别采用含 10%胎牛血清、8μg/L表皮生长因子(EGF)、10%胎牛血清+8μg/L EGF 、10%胎牛血清+8μg/L EGF+30%成纤维细胞培养 基、10%胎牛血清+30%成纤维细胞培养基的DMEM培养液进行培养，于培养前及培养后7 、14d观察各组细胞的光镜特 征，采用免疫组织化学染色检测特异性细胞表面标记物的表达以及Western blotting 方法检测诱导前后ADSCs 中CK-7 、 CK-14 的蛋白表达水平。结果　与未加EGF诱导剂培养组比较，经EGF诱导14d后多数ADSCs 细胞形态、排列方式发生 改变。CK-19免疫组化染色显示呈强阳性。Western blotting 检测结果提示诱导后的黏膜上皮细胞特异性蛋白CK7 、CK14 呈高表达。结论　ADSCs 在一定诱导条件下可向黏膜上皮样细胞分化。 [关键词]　脂肪干细胞；表皮生长因子；上皮样细胞；细胞分化 [中图分类号]　Q254　　　　[文献标志码]　A　　　　[文章编号]　0577-7402(2015)10-0798-05 [DOI]　10.11855/j.issn.0577-7402.2015.10.06 In vitro study on differentiation of adipose-derived stem cells into mucosal epithelial cells * WANG Cheng, XU Xiao, CHEN Chong, LI Liang, YAN Jun-ling, TANG Su-yang Department of Skin and Hair Regenerative Medicine, General Hospital of Chinese People’s A rmed Police Forces, Beijing 100039, China * Corresponding author, E-mail: 2278645614@ [Abstract]　Objective　To explore the possibility of inducing the differentiation of adipose-derived stem cells (ADSCs) into mucosal epithelial cells in vitro. Methods　Adult subcutaneous adipose tissue was obtained for carrying out the isolation, culture, amplification and differentiation of ADSCs, and to identify the genetic stability of the third passage. The cultures were performed with DMEM culture media containing respectively 10% fetal bovine serum (FBS), 8 μg/L epidermal growth factor (EGF), 10% FBS+8 μg/L EGF, 10% FBS+8 μg/L EGF+30% fibroblast cell culture medium, and 10% FBS+30% fibroblast culture medium. The light microscopic characteristics of the cells were observed before culture and 7 and 14 days after culture. The expression of specific cell surface markers and