Objective：This study aims to establish a co-culture system of matured stromal vascular fraction （SVF） with Hepa1-6 and study the effects on lipid metabolism of hepatocytes. Method：Primary SVF cells were isolated from mice subcutaneous fat and induced to matured SVF cells with differentiation medium. Matured SVF cells 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 PPARγ and PGC-1α in SVF cells and lipid metabolism related genes such as CD36，FATP2 and GPAT1 in Hepa1-6. Cellular triglyceride level of co-cultured Hepa1-6 was detected by GPO Enzyme Method. Results：Isolated primary SVF cells 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 PPARγ and PGC-1α of SVF cells was significantly increased. The cellular triglyceride 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 SVF cells with Hepa1-6 in Transwell insert has been successfully established. Co-culture with mature SVF cells induces lipid accumulation in Hepa1-6. This process may be maintained by the elevated expression level of CD36 and GPAT1，increasing fatty acid absorption and triglyceride synthesis.