In vitro mesenchymal stem cells differentiation into hepatocyte-like cells in the presence and absence of 3D microenvironment

Abstract

Our study was aimed to select the best growth factor cocktail for differentiation of mesenchymal stem cells (MSCs) into hepatocytes-like cells (HLCs) in absence and presence of three dimensional (3D) microenvironment. Ninety milliliters of bone marrow was aspirated from the iliac bone for separation of MSCs. This study was conducted on 20 patients with advanced liver cirrhosis. They were divided into two groups. Group I; MSCs were plated onto 96-well plates without microenvironment (control group). Group II; MSCs were plated onto 96-well plates with 3D microenvironment. Surface expression of CD271, CD29, and CD34 were analyzed using flow cytometry. MSCs were differentiated in vitro into HLCs by adding four growth factor cocktails in presence and absence of 3D microenvironment. Hepatogenesis was assessed by immunohistochemical analysis of OV6, ?-fetoprotein, albumin, and cytokeratin 18 expressions. There was statistically significant increase in the expression of CD271 and CD29 after MSCs culture (P < 0.001). Heterogeneous cell population composed of MSCs and differentiated HLCs were encountered (40 % hepatocytes and 60 % MSCs). Furthermore, our results showed that growth factor cocktails 1 and 2 gave the best result for differentiation of MSCs into HLCs. MSCs could be feasible, readily available, and novel source for differentiation of MSCs into HLCs for future therapeutic implication.