Unrestricted somatic stem cells differentiation into hepatic-like cells in a three dimensional arrangement

Abstract

Reports describing the human cord blood derived mesenchymal stem cells (hUSSCs) hepatocytic differentiation potential are rare and there is no report describing fibrin base 3D culture of hepatocyte-like cells differentiated from cord blood unrestricted somatic stem cells. Hepatocyte-like differentiated stem cells has the potential to provide compensation for acute liver failure patients. Therefore, we characterized cord blood Unrestricted Somatic Stem Cells (USSC) and their hepatocyte differentiation capabilities for functional liver support in a three dimensional arrangement. Moreover, three-dimensional (3D) network and its distribution to grow and differentiate stem cells in so far as possible mimic to native tissue holds huge potential in liver tissue engineering. In this study, we isolate USSCs from human cord blood and validate them with flow-cytometry. Briefly, we differentiated cells using DMEM containing FGF4, Activin A, HGF and 2% FBS into hepatocyte-like cells (HLCs) in two conditions; 3D state in biocompatible fibrin Hydrogel and or conventional culture (2D) in polystyrene plates for 21 days. Immuncytochemical and gene expression analysis revealed the expression of KRT18, KRT19, albumin, Sox17, FoxA2 and Prox1 increased in 3D fibrin culture compared to 2D conventional culture. In combination, these data reveal that using 3D networks can resemble near natural tissue properties for effective generating HLCs which used to cell replacement in the future. As well as, USSC represent a stem cell source with a substantial hepatic differentiation capability which hold the possible for clinical applications.