体外诱导脂肪来源间充质干细胞间质-上皮转换

Induction of mesenchymal-epithelial transition of human adipose-derived mesenchymal stem cells in vitro

目的探讨在体外诱导人脂肪来源间充质干细胞(hAD-MSCs)发生间质-上皮转换(MET)的可行性。方法从人脂肪中分离、培养间充质干细胞,流式细胞术鉴定其细胞表面标志。在体外添加激活素A(activin A)、维甲酸(RA)和骨形态发生蛋白7(BMP7)培养后,倒置显微镜下观察hAD-MSCs诱导后的形态变化,采用RT-PCR方法检测间质和上皮细胞相关基因vimentin,E-cadherin,ZO-1和β-catenin在诱导分化过程中表达水平改变。Western blot检测诱导前后间质标记vimentin和上皮标记β-catenin的表达。结果诱导后细胞形态发生改变,由长梭形变为卵圆形,上皮标记基因E-cadherin,ZO-1和β-catenin的mRNA表达显著上调(P<0.05),同时间质标志基因vimentin的mRNA表达显著下降(P<0.05)。Western blot结果显示,与未诱导组相比,诱导后细胞开始表达β-catenin,而vimentin的表达明显下降(P<0.05)。结论 Activin A、RA和BMP7联合使用可诱导hAD-MSCs发生MET的过程。

Objective To explore the feasibility of the induction of mesenchymal-epithelial transition of human adi- pose-derived mesenchymal stem ceils in vitro. Methods MSCs were isolated from human adipose and induced with activin A, RA, BMP7 in vitro. The morphological changes were observed and the expressing of protein vimentin and β-catenin were analysed by Western blot. The mRNA expression level of vimentin, E-cadherin, ZO-1 and 13-catenin was compared in induced and non-induced groups by semi-quantitative reverse transcription polymerase chain reaction （RT-PCR）. Results After inducing, hAD-MSCs morphology changed from spindle to orbicular-ovate, and expressed epithelial identification marker. The mRNA expression levels of E-cadherin, ZO-1 and 13-catenin were significantly higher （P 〈 0. 05 ）, vimentin was lower in induced group than those in non-induced group （ P 〈 0. 05 ）. Conclusions Human adipose-derived mesenchymal stem cells can be induced into epithelial-like cells in vitro.