体外诱导脂肪来源干细胞向黏膜上皮细胞分化的研究  

In vitro study on differentiation of adipose-derived stem cells into mucosal epithelial cells

作　　者：王程[1] 徐潇[1] 陈冲[1] 李梁[1] 闫俊灵[1] 汤苏阳[1] 

机构地区：[1]北京武警总医院皮肤再生医学科,100039

出　　处：《解放军医学杂志》2015年第10期798-802,共5页Medical Journal of Chinese People's Liberation Army

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。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）, 8μg/L epidermal growth factor （EGF）, 10% FBS＋8μg/L EGF, 10% FBS＋8μg/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 expression levels of CK-7 and CK-14 proteins in ADSCs before and after induction were detected by immunohistochemical staining and Western blotting respectively. Results Compared with the group without EGF, the morphology and arrangement of ADSCs induced by EGF for 14d were changed. Immunochemical staining showed CK-19 was strongly positive expressed. Western blotting analysis showed that CK-7 and CK-14 were strongly expressed in the epithelial cells. Conclusion Adipose-derived stem cells can be differentiated into mucosal epithelioid cells in vitro.

关 键 词：脂肪干细胞 表皮生长因子 上皮样细胞 细胞分化 

分 类 号：Q254[生物学—细胞生物学]