Hang Hualian, Yu Yabin, Wu Ning, Huang Qingfeng, Xia Qiang, Bian Jianmin
Department of Liver Surgery, Ren Ji Hospital, School of Medicine, Jiao Tong University, Shanghai, China; Department of General Surgery, Nanjing Hospital Affiliated to NanJing Medical University, Nanjing, China.
Department of General Surgery, Nanjing Hospital Affiliated to NanJing Medical University, Nanjing, China.
PLoS One. 2014 Aug 19;9(8):e104133. doi: 10.1371/journal.pone.0104133. eCollection 2014.
To investigate the differentiation potential of human umbilical mesenchymal stem cells (HuMSCs) and the key factors that facilitate hepatic differentiation.
HuMSCs were induced to become hepatocyte-like cells according to a previously published protocol. The differentiation status of the hepatocyte-like cells was examined by observing the morphological changes under an inverted microscope and by immunofluorescence analysis. Hepatocyte nuclear factor 4 alpha (HNF4α) overexpression was achieved by plasmid transfection of the hepatocyte-like cells. The expression of proteins and genes of interest was then examined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) or real-time RT-PCR methods.
Our results demonstrated that HuMSCs can easily be induced into hepatocyte-like cells using a published differentiation protocol. The overexpression of HNF4α in the induced HuMSCs significantly enhanced the expression levels of hepatic-specific proteins and genes. HNF4α overexpression may be associated with liver-enriched transcription factor networks and the Wnt/β-Catenin pathway.
The overexpression of HNF4α improves the hepatic differentiation of HuMSCs and is a simple way to improve cellular sources for clinical applications.
研究人脐间充质干细胞(HuMSCs)的分化潜能以及促进肝分化的关键因素。
根据先前发表的方案将HuMSCs诱导成为类肝细胞。通过在倒置显微镜下观察形态变化以及免疫荧光分析来检测类肝细胞的分化状态。通过对类肝细胞进行质粒转染实现肝细胞核因子4α(HNF4α)的过表达。然后通过蛋白质印迹法以及逆转录-聚合酶链反应(RT-PCR)或实时RT-PCR方法检测感兴趣的蛋白质和基因的表达。
我们的结果表明,使用已发表的分化方案可轻松将HuMSCs诱导成为类肝细胞。诱导的HuMSCs中HNF4α的过表达显著提高了肝脏特异性蛋白质和基因的表达水平。HNF4α过表达可能与肝脏富集转录因子网络和Wnt/β-连环蛋白通路有关。
HNF4α的过表达改善了HuMSCs的肝分化,是改善临床应用细胞来源的一种简单方法。