The different methods to induce the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells

Wang Hui; Zhang Jian-jun

doi:10.3969/j.issn.2095-4344.2014.41.009

Abstract：BACKGROUND:At present, heterologous serum as a medium is a common method for culture of bone marrow mesenchymal stem cells, but it is limited by the disease transmission between species, the potential immune rejection and even controversial ethic issues. This method is also contrary to the requirements of the Ministry of Health. Autologous platelet-rich plasma is a kind of whole blood extract, containing a variety of growth factors. OBJECTIVE:To explore the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells cultured in autologous platelet rich plasma alternative to traditional heterogeneous serum-free culture. METHODS:8 mL bone marrow from the rabbit iliac crest was extracted and anti-coagulated with heparin, and then bone marrow mesenchymal stem cells were isolated using density gradient centrifugation. The cells were divided into autologous platelet rich plasma group (10%autologous platelet rich plasma) and fetal bovine serum group (10%fetal bovine serum). At the passage 4, the cells in the two groups were respectively subdivided into experimental and control groups. Experimental groups were subjected to osteogenic induction, while no change was done in the control groups. cellproliferation was determined by using growth curves;the activity of alkaline phosphatase was detected to adjust the osteogenic differentiation of cells in different groups. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells at different generations al showed good proliferation. At 12 days of autologous platelet rich plasma culture, the activity of alkaline phosphatase in the passage 4 cells in the experimental group was significantly higher than that in the control group;while the activity of alkaline phosphatase in the autologous platelet rich plasma group was significantly higher than that in the fetal bovine serum group (P<0.01). These findings indicate that autologous platelet rich plasma as a substitute of xenogeneic serum for culture of bone marrow mesenchymal stem cells is a method characterized as safe and reliable, simple operation, high-purity active induction.