Comparison and Optimization: Different Medium and a Novel Scheme for Rabbit Bone Marrow Mesenchymal Stem Cells Culture

Abstract

BackgroundRabbit bone marrow mesenchymal stem cells (rBMSCs) are widely used as seed cells for bone tissue engineering, but the effect of different medium on the physiological characteristics of rBMSCs is still not clear and the optimal culture medium for rBMSCs is also uncertain.MethodsRBMSCs were divided into 4 groups: DMEM/F12 (F12) group, DMEM/Low glucose (LG) group, DMEM/High glucose (HG) group, and optimizational culture method group which was cultured with DMEM/High glucose for the first 6 days and then DMEM/Low glucose (HG-LG). The morphology, proliferation ability, stem cell surface markers, multi-directional differentiation potential, adhesion ability and vitality in tissue engineering bone (TEB) were detected and compared. ResultsRBMSCs in the 4 groups shared the same shape, similar surface markers (positive in CD44, CD29, negative in CD45, CD34) and similar osteogenic, adipogenic and chondrogenic directional differentiation capability. For the primary rBMSCs, the proliferation ability of HG group was higher than that of LG group (P<0.05), while F12 group was between the former two. The proliferation ability of the 3rd passage of rBMSCs in the 4 groups was similar. The adhesion ability of the 3rd passage rBMSCs in the 4 groups was listed as follows starting from high to low: LG group and HG-LG group (they had similar ability), F12 group, HG group, and the difference was statistically significant (P<0.05). The vitality of TEB in LG group and HG-LG group was higher than that in F12 group and HG group (P<0.05). ConclusionsThe primary rBMSCs cultured with DMEM/High glucose showed good proliferation ability, while the 3rd rBMSCs cultured with DMEM/Low glucose showed good adhesion ability and vitality in TEB. RBMSCs cultured with DMEM/High glucose for the first 6 days and then DMEM/Low glucose showed good proliferation ability, good adhesion ability and good vitality in TEB, which might provide a novel optimizational scheme for rBMSCs culture.