Objective:To observe the growth and function of primary mouse islets in the liver decellularized bioscaffold(LDB),and explore the new method of tissue engineering for treatment of diabetes. Methods:We perfused the whole mouse liver using detergent through hepatic portal vein in a continuous way and manufactured the whole LDB with the complete structure. The collagenase P was perfused into the mouse common bile duct,and the number of the isolated isles was counted and the integrity of the islets’ structure and function were analyzed. The isolated primary mouse islets were transplanted into the LDB and cultured for 5 days in the three-dimensional culture system. Then we appled HE staining,insulin immunohistochemical analysis and fluorescence quantitative polymerase chain reaction(PCR). Results:The LDB maintained their original round shape after decellularization and demonstrated a complete lack of nuclear staining and blood vessel. Collagenase 1 fluorescent staining showed that the collagen structure intact. The DNA quantification showed(38±11)ng/mg dsDNA in dry weight of ECM. In vivo biocompatibility test of the LDB showed that there didn’t exist any pathological presentation. The result of dithizone(DTZ)staining showed that the staining islets had the scarlet color Glucose stimulation test showed that high quality islets results in a greater insulin stimulation at high glucose concentration than that observed at low glucose concentration (P < 0.01). Insulin immunohistochemistry examination showed that the LDB parenchyma in transplanted area had brown stain. Insulin gene expression displayed the expression level of insulin in LDB was two times greater than the level of plate culture with statistical significance(P < 0.01). Conclusion:In LDB 3D culture system,islets showed better cell viability and more effective insulin secretion function than traditional plate culture.