Isolation, Cultivation and Biological Features of Adult Bone Marrow Mesenchymal Stem Cells in vitro

Objective:To establish a method for isolation, cultivation and expansion of adult bone marrow mesenchymal stem cells in vitro,and study their some biological features and a method for inducing their differentiation into adipocytes,which could be applied to identify what we cultured.Methods:MSCs were isolated from adult bone marrow by Percoll, cultured on fibronectin with epidermal growth factor and platelet-derived growth factor BB and 2% fetal bovine serum,and purified by their adhesiveness; the changes in cell morphology and the ultrastucture were observed,and cell surface markers were detected by Flow cytometry;MSCs were induced to differentiate into adipocyes by change to 10% fetal bovine serum and nicotinamide.Results:Both primary and passage MSCs have maintained highly proliferative capacity. The ultrastructure showed the young cell nature of stem cell.The cells were negative for CD34, CD45, CD19, HLA-DR,but positive for CD44,CD10, CD29,CD13.MSCs can be induced to differentiate into adipocytes that stained with Oil-red O.Conclusion:With our method, MSCs can be cultured stably,and expanded quickly; they can be induced to differentiate into adipocytes under given cultured conditions in vitro;and the cells we cultured have the same characters as MSCs.