In vitro cultivation, identification and osteoinduction of adult bone marrow mesenchymal stem cells

Zhang Ming-jie; Zhang Qing-wen; He Wei; Chen Zhen-qiu; Ou Zhi-xue; Jia Xiao-jun

doi:10.3969/j.issn.2095-4344.2013.45.019

Abstract：BACKGROUND:Bone marrow mesenchymal stem cells have the potential of self-proliferation and multi-directional differentiation, while mesenchymal stem cells are few in adult bone marrow. In vitro purification, amplification and osteoinduction are very important for the research of bone tissue engineering. OBJECTIVE:To establish a simple and reliable in vitro cultivation and identification system of adult bone marrow mesenchymal stem cells, and to induce the mesenchymal stem cells to differentiate into osteoblasts. METHODS:Bone marrow were extracted from adult anterior superior iliac, the density gradient centrifugation and adhesion method were used to isolate, purify, culture and amplify the bone marrow mesenchymal stem cells. Osteogenic medium was prepared by mixing appropriate amount of dexamethasone,β-glycerophosphate and ascorbic acid C. The cells were divided into osteoinduction group and blank control group for observation. <br> RESULTS AND CONCLUSION:Adult bone marrow mesenchymal stem cells were in typical long spindle-shape. The cells grew into rapid proliferation phase at 8-11 days and the growth curve was S-shape. CD44 and CD90 were in positive expression, while CD34 and CD45 were negative. The alkaline phosphatase activity was increased with culturing time prolonging, and reached the summit at the 12th day. The alkaline phosphatase activities of osteoinduction group were higher than those in the blank control group at different time points. These results suggested that in vitro cultivation, identification and osteoinduction system could obtain mesenchymal stem cells with high purity and good osteogenic differentiation capacity.