Isolation,Culture in vitro,Identification and Induction into Adipocytes of Rabbit Bone Marrow Mesenchymal Stem Cells

Objective To isolate,culture in vitro and identify the rabbit bone marrow mesenchymal stem cells(MSCs mesenchymal stem cells) and induce them to differentiate into adipocytes.Methods Health juvenile Japan big-eared white rabbits were used,and the bone marrow was collected after rabbits were euthanized by anesthesia.MSCs were isolated and cultured by direct adherent culture,then the morphology of MSCs was observed.At the same time,MSCs were indentified by detecting the four surface antigens including CD34,CD44, CD45 and CD90 through flow cytometry.The fifth generation of MSCs were induced to differentiate by addition of 50 μmol/L ascorbate2phosphate,50 μmol/L indomethacin and 0.5umol/Ldexamethasone.Results Flow cytometry showed CD34(-),CD4(-),CD44(+) and CD90(+) in the second generation of MSCs,which were in accordance with the surface antigens of MSCs.After the fifth generation of MSCs were induced for ten days to differentiate into adipocytes,oil red O staining showed that a large number of lipid deposition and mutual confluence,and the shape of cells became polygon from long shuttle.Conclusions The cells cultured in vitro by our method are pure mesenchymal stem cells,have the typical characteristics of stem cells,and can differentiate into adipocytes.The mesenchymal stem cells cultured in vitro can be used to study the cell differentiation mechanisms in tissue engineering and animal models.