حسین رضوان

Background Isolation and culture of bone marrow mesenchymal stem cells is the first step in all stem cell studies, although isolation of these cells from BALB/c mice bone marrow is not as easy as other animals. Hence, developing an efficient method with high simplicity for isolation of bone marrow stem cells has always been a goal for stem cell researchers. Materials and Methods After anesthetizing Balb/c mice, femur and tibia bones were removed and the bone marrow was flashed out with DMEM cell culture medium containing Penicillin/Streptomycin (100u/ml)/(0/1mg/ml) using a 10ml siring. The cells were then harvested and cultured in T25 flasks containing 5ml of the cellmedium. The cell medium was replaced every 24 hours for 3 times and the cells were recultured at three days interval for three times. To confirm the stem cell identity, the cells were induced to be differentiated into osteocytesand adipocyts by culturing the cells in differential medium. The expression of stem cell markers and the lack of hematopoietic markers on the cells were also investigated. Results The cells were isolated from the mouse bone marrow and differentiated to adipocyts or osteocytescell lines. The expression of CD90, CD44, CD73 and CD105 on the mesenchymal stem cells are 74%, 79%, 86% and 81%, respectively. The expression of CD34 and CD45 on the cells were also 8.7% and 0.28%, respectively. Conclusion The results showed that BALB/c mice bone marrow stem cells can be efficiently isolated and cultured by this method.