Abstract: Objective: To study differentiation of human bone marrow derived mesenchymal stem cells （BDMSC） into heoatocytes for ideal cell origin of liver tissue engineering. Methods: After vary days of induction with hepatocyte growth factor（HGF） and fibroblast growth factor 4 （FGF-4） in 3D Matrigel, BDMSC and heoatocyte-like cells were determined by the utilization of morphological observation, immunofluorescence of alpha-fetoprotein（AFP） and albumin（Alb）and uptake and excretion of Indocyanine green（ICG） that were special for heoatocytes. Results: After 3D-cultured and induced with HGF and FGF-4 in vitro in Matrigel for 3-7 days, BDMSCs came into heoatocyte-like cells that expressed AFP and Alb and possessed ability of uptake and excretion of ICG. Conclusion: In vitro HGF、FGF and Matrigel maybe supply a proper microenviroment and play an important role during process of heoatocytes differentiation of BDMSC. This study offered a theoretical and technical foundation of mesenchymal stem cells （MSC） application for liver tissue engineering.