hE-Cadherin-Fc protein engineered biomaterial enhancing the growth of human mesenchymal stem cells

摘要

Biomaterials can play important roles as extracellular matrix in tissue engineering and regenerative medicine by controlling cellular behaviors and functions. In recent years, in order to biomimic construct the extracellular microenvironment, biosynthesis materials by gene engineering used in artificial ECMs had gained special interest because their selectivity to specific lineages of cells or tissues.. In this study, we design an artificial ECM with E-Cadehrin extracellular domain to control the proliferation and differentiation of human mesenchymal stem cells. The hE-Cadherin-Fc fusion protein was prepared by the FreeStyle MAX expression Systems with recombinant plasmid gene composing extracellular domain of hE-Cadherin and Fc domain of IgG and then purified by Protein A affinity chromatography. The fusion protein was immobilized on the hydrophobic material of polystyrene and then was used as the artificial ECM for culturing human bone marrow mesenchymal stem cells (hMSCs). Comparing with that growth on the surfaces of polystyrene, a higher cell adhesion and more spread cellular morphologies were observied by FITC-phalloidin/PI staining and a higher proliferation of hMSCs was also shown by MTT assay and FDA/PI staining on the hE-cadheirn-Fc modified surfaces. Furthermore, the expressions of hepatic markers (Albumin, AFP, HNF4, CK18 and CYP3A4) were detected by real time PCR and flow cytometry (CD117 and CD105) respectively, showing that E-Cadherin-Fc immobilized surfaces enhanced the inducing efficiency of hMSCs differentiation to hepatocyte-like cells under condition medium with appropriate induced factors (Dex, OSM, HGF and FGF4). We suggest that gene engineered biomaterial of hE-Cadherin-Fc fusion protein should be an excellent artificial ECM for enhancing hMSCs proliferation and hepatic differentiation.