Objective: CYP450 is considered as one of the most vital metabolizing enzymes in hepatic detoxification, secreted by the functional liver cells, termed hepatocytes. Several aspects of hepatocyte-like cells derived from mesenchymal stem cells have been studied; however, less attention has been paid to the drug metabolizing ability of these in vitro differentiated cells. The aim of this study was to evaluate the effect of laminin coating on CYP450 enzyme activity in our differentiated BM-MSCs. Materials and Methods: The characteristics of MSCs obtained from Royan were confirmed by immunophenotyping analysis and their differentiation potential into osteocytes and adipocytes. Hepatogenic differentiation was induced by hepatocyte growth factor, oncostatin M and dexamethasone. On day 21, “P450 CYP3A4 screening system” was used to study the enzymativ activity. This method was a luminescence assay in which a CYP substrate converted by CYP enzymes to a luciferin product that in turn reacted with a Luciferin Detection Reagent to produce light. The amount of light produced was proportional to the amount of luciferin produced after the CYP reaction. Undifferentiated human MSCs were used as negative control. Results: The flowcytometry findings revealed that over 90% of the cells expressed the MSC markers and not hematopoietic and leukocyte markers. The results showed that the activity of CYP3A4 was two times higher in the cells differentiated on laminin-coated vessels Compared to that of the Polystyrene and the differences were Significant statistically (p < 0.05). Increasing activity of this enzyme showed improved differentiation in laminin matrix. Furthermore, the results of P450 CYP3A4 screening system demonstrated that this enzyme had no activity in the human BM-MSCs before differentiation. Conclusion: Together, these findings may indicate that laminin coating can improve the differentiation of hepatocyte- like cells from human BM-MSCs, at least in part by stimulating their CYP450 enzyme activity, which is fundamental for their drug detoxification function.