Objective To provide experimental data for preclinical studies on the treatment of human diseases byautologous fat stem cell transplantation, including isolation and culture methods and the biological characteristics oflaboratory animals including C57 mice, SD rats, and cynomolgus monkey adipose-derived stem cells (ADSCs), which werecompared with those of humans. Methods hADSCs, mouse adipose derived mesenchymal stem cells (mADSCs), ratadipose derived mesenchymal stem cells (rADSCs) and cynomolgus monkey adipose derived mesenchymal stem cells(cADSCs) were identified via surface markers using flow cytometry, and adipogenic and osteogenic differentiation wasinduced. Results All the hADSCs, mADSCs, rADSCs and cADSCs showed fibroblast-like morphology. Flow cytometryshowed that mesenchymal stem cell surface markers CD90 and CD29 in ADSCs of each species were positive, while thevascular endothelial cell marker CD31 was negative. Oil red O and alizarin red staining showed that hADSCs, mADSCs,rADSCs and cADSCs differentiated into fat cells and bone cells. Conclusions Laboratory animal ADSCs and hADSCs canbe cultured using the same method and they all had similar surface marker expression, as well as adipogenic and osteogenic differentiation potential, indicating their usefulness for the pre-clinical study of autologous ADSC transplantation.