Isolation,Culture and Basic Biological Properties of Rat Bone Marrow Mesenchymal Stem Cells in Vitro

Objective:To seek the way of isolating and culturing rat bone marrow mesenchymal stem cells(MSCs), and to investigate the basic biological characteristics of rat bone marrow MSCs. Methods: Bone marrow was collected from femur of 5～8 weeks old female rats and seeded in cell culture flask. the morphology of MSCs after passage 5 were analyzed. The growth curve and cytochemistry were observed. The surface antigens and cell cycle were analysed by flow cytometer(FCM). Results:MSCs had spindle and polygon shape . The growth curve likes S shape. The double time of MSCs was about 30 hours. Cytochemistry evaluation showed that MSCs were homogeneously negative for ALP, POX, SB and positive for α-NAE, PAS. MSCs expressed CD29,CD44,CD90 and didn't express CD34,CD38,CD45.Cell cycle included G 0/G 1 phase:79.48%,G 2/M phase: 6.10%,S phase 14.42%. Conclusion: The MSCs from rat bone marrow showed stable, long-term growth and proliferated quickly. It can be served as optimal cell source for tissue engineering and for transgenic research.