Isolation,Characterization and Differentiation of Rabbits Adipose-derived Stem Cells

Objective: To isolate and culture rabbits adipose-derived stem cells(ASCs),and to observe the biological characteristics of rabbits ASCs and to analyze the multipotent properties of differentiation with proper condition in vitro.Method: ASCs were prepared by collagenase Ⅰ digestion of the inguinal fat pads from rabbits,and cultured at 37℃ in a 5% CO2 humidified incubator.Cell passage was performed following about 80%-90% confluence.The growth curve was drawn.And Hematoxylin-eosin(HE) staining was used to observe the morphology of ASCs.The surface antigens of ASCs were analyzed by flow cytometry.Passage 3 of ASCs were used to differentiated into adipocytes,osteoblasts,and identified by the Oil Red O staining and Alizarin red staining.Results: Adherent ASCs showed fibroblast-like and spindle shape or polygon shape,strong proliferative ability and active differentiation.The growth curve was like 'S' shape.Flow cytometry showed that ASCs expressed CD29,but did not express CD31.The Oil Red O staining and Alizarin red staining showed positive reaction.