Acquisition and biological characteristics of rabbit bone marrow derived mesenchymal stem cells

Objective:To cultivate rabbit bone marrow derived mesenchymal stem cells(MSCs)in vitro,and find a set of amplification system,then observe its biological characteristics.Methods:The MSCs from rabbit bone marrow were isolated,purified and amplified by density gradient centrifugation and adherent culture in vitro.The changes of MSCs morphology,growth and proliferation were observed by inverted phase contrast microscope,and MSCs ultrastructure and cell cycle were detected by transmission electron microscope and flow cytometer respectively;MSCs were induced into osteogenetic cells or adipose cells and then were verified.Results:The primary cells and the passage cells were mostly fusiform in shape,which was similar to fibroblast cell;during adherence growth,they were arranged tightly in a swirling pattern;the cell cycle analysis showed that more than 87% cells were in G0/G1 phase;after MSCs were induced,cell alkaline phosphatase stain showed that alkaline phosphatase was positive in osteogenetic cell and formed mineralization nodus;oil red O stain showed that red lipid droplet existed in adipose cell.Conclusion:MSCs can be isolated and cultivated by the method of density gradient centrifugation and adherent culture in vitro,the potentiality of proliferation and multi-directional differentiation can meet the requirements of the further experiments.