Introduction: Efforts have been taken to find appropriate sources to replace liver transplantation. Wharton’s jelly is an unlimited source of stem cells that can be used in cell therapy and tissue engineering. In this study we investigated whether Wharton’s jelly-derived mesenchymal stem cells (WJMSCs) could trans-differentiate into hepatocyte in the presence of mouse liver cell-free extract. Materials and Methods: Mesenchymal stem cells (MSCs) were isolated from the umbilical cord. The cells were permeabilized by Sterptolysin O in the presence of mouse liver cell-free extract for 21 days. To evaluate differentiation and morphological changes, immunostaining for cytokeratin 18 and 19 were performed for differentiated and control cells. Functional assays were done by periodic acid Schiff (PAS) stain. Results: The phenotype of treated MSCs in the presence of liver cell-free extract changed into polygonal cells. Immunostaining demonstrated the expression of cytokeratin 18 and 19 in differentiated cells. Glycogen storage in differentiated cells revealed by PAS staining indicated functional property of differentiated cells. Conclusion: It seems that factors existing in the extract able to trans-differentiate WJMSCs into functional hepatocyte.