Abstract：Objective: To establish a co-culture system of matured stromal vascular fraction with Hepa1-6 and study the effects on lipid metabolism of Hepatocytes. Method: Primary SVFs were isolated from mice subcutaneous fat and induced to matured SVFs with differentiation medium. Matured SVFs were co-cultured with Hepa1-6 by using Transwell system for 2 days. RT-PCR was applied to detect the expression of differentiation biomarker such as PPAR? and PGC-1α in SVFs and lipid metabolism related genes such as CD36, FATP2 and GPAT1 in Hepa1-6. Cellular TAG level of co-cultured Hepa1-6 was detected by GPO Enzyme Method. Results: Isolated primary SVFs were successfully differentiated to mature SVF cells with significantly large lipid droplets after treated with adipocytes differentiation medium for 6 days. The expression level of PPAR??and? PGC-1α of SVFs was significantly increased. The cellular TAG level of co-cultured Hepa1-6 was significantly higher than that of control group accompanied with increased expression level of GPAT1 and CD36. Conclusion: The co-cultured system of matured SVFs with Hepa1-6 in Transwell insert has been successfully established. Co-culture with mature SVFs induces lipid accumulation in Hepa1-6. This process may be maintained by the elevated expression level of CD36 and GPAT1.