人E钙粘素融合蛋白基质对人骨髓间充质干细胞向肝细胞定向诱导分化的影响研究

Human E-Cadherin-Fc Fusion Protein Matrix Improving the Hepatic Differentiation of Human Mesenchymal Stem Cells

人骨髓间充质干细胞(hBMSCs)源的肝细胞对于肝脏疾病的治疗和药物的开发具有巨大的应用潜力。利用基因工程技术构建了融合蛋白质粒,并通过真核细胞表达体系表达出了人E-钙粘素胞外域与免疫球蛋白Fc段的融合蛋白(hE-cadherin-Fc),并将其用于疏水材料的表面修饰仿生构建细胞-细胞间相互作用的细胞外微环境,检测其对hBMSCs向肝样细胞定向诱导分化的影响。诱导分化培养4周后,与组织培养板(TC-PS)、明胶(Gelatin)基质相比,hE-cadherin-Fc基质显著促进细胞表达ALB、CK18、HNF-4等肝细胞分化基因,并且细胞的糖原合成和吲哚青绿(ICG)摄取功能均显著提高。在hE-cadherin-Fc基质表面分化培养4周时,白蛋白和尿素合成的量为2.7pg/细胞/d和36.7 pg/细胞/d,相对于TC-PS和Gelatin分别提高了42.1%和28.6%、57.5%和25.7%。综上所述,利用hE-cadherin-Fc基质化构建细胞外微环境,有利于促进人骨髓间充质干细胞向功能化肝细胞的定向分化。

Hepatocytes differentiating from human mesenehymal stem cells （hBMSCs） hold great potentials for cell-based approaches to liver diseases and drug development. In order to effectively expand and induce hepatic differentiation of hBMSCs, a fusion protein consisting of human E-cadherin extracellular domain and the immunoglohulin G Fc region （hE-cadherin-Fc） was biosynthesized and used as an extracellular matrix biomimicking epithelia cell adhesion junction. The effects of the hE-cadherin-Fc matrix on the hepatic differentiation of hBMSCs were studied in this paper. Compared with TC-PS and gelatin coated surfaces, the hE-cadherin-Fc matrix effectively stimulated the hBMSCs to progress toward the polygonal morphology of hepatocyte, and enhanced the differentiated ceils expressing hepatocyte-specific genes and live-specific functions. These results show hE-cadherin-Fc is a promising artificial extracellular matrix （ECM） for the hBMSCs differentiation via homophilic interaction of hE-cadherin and synergy between cell adhesion molecular and growth factors.