Laminin matrix promotes hepatogenic terminal differentiation of human bone marrow mesenchymal stem cells

Abstract

<strong><em>Objective(s)</em><em>:</em></strong>The application of stem cells holds great promises in cell transplants. Considering the lack of optimal <em>in vitro</em> model for hepatogenic differentiation, this study was designed to examine the effects of laminin matrix on the improvement of <em>in vitro</em> differentiation of human bone marrow mesenchymal stem cells (hBM-MSC) into the more functional hepatocyte-like cells. <strong><em>Materials and Methods</em><em>:</em></strong>Characterization of the hBM-MSCs was performed by immunophenotyping and their differentiation into the mesenchymal-derived lineage. Then, cells were seeded on the laminin-coated or tissue culture polystyrene (TCPS). The differentiation was carried out during two steps. Afterward, the expression of hepatocyte markers such as AFP, ALB, CK-18, and CK-19 as well as the expression of C-MET, the secretion of urea, and the activity of CYP3A4 enzyme were determined. Moreover, the cytoplasmic glycogen storage was examined by periodic acid–Schiff (PAS) staining. <strong><em>Results:</em></strong>The results demonstrated that the culture of hBM-MSC on laminin considerably improved hepatogenic differentiation compared to TCP group. A significant elevated level of urea biosynthesis and CYP3A4 enzyme activity was observed in the media of the laminin-coated differentiated cells (<em>P</em><0.05). Furthermore higher expressions of both AFP and ALB were determined in cells differentiated on laminin matrix. Glycogen accumulation was not detected in the undifferentiated hBM-MSCs, however, both differentiated cells in laminin and TCPS groups demonstrated the intracellular glycogen accumulation on day 21 of hepatogenic differentiation. <strong><em>Conclusion:</em></strong>Taken together, these findings may indicate that laminin matrix can improve terminal differentiation of hepatocyte-like cells from hBM-MSCs. Thus, laminin might be considered as a suitable coating in hepatic tissue engineering designs.