Isolation, Culture, Differentiation and Identification of Mice Adipose-derived Stem Cells

Objective: By isolating and cultruring adipose-derived stem cells(ASCs) of C57 mice,to observe the biological characteristics and explore multipotent differentiation properties of ASCs in vitro.Methods: Cells were isolated from the bilateral inguinal fat pads from C57 mice by 0.25% collagenase I digestion,cultured at 37 ℃ with 5% CO2 humidi fl ed incubator and expanded after reaching 80% confluence.Cells morphology were observed.The growth curve were drawn by MTT colorimetry.The surface markers of cells were analyzed by flow cytometry.Passage 2 cells were cultured in osteogenic or adipogenic medium for 3 weeks respectively,and verified with Alizarin red staining or Oil Red O staining.Results: ASCs presented fusiform shape and fibroblast-like,The growth curve was like 'S' shape,proved strong proliferative ability of ASCs.The results of Flow cytometry showed that ASCs expressed CD29,CD44 and CD90,but not CD31,CD 4 and CD45.Alizarin red staining and Oil Red O staining showed positive reaction.Conclusions: The cells were substantiated as ASCs,being of precise differentiation potentiality.