Isolation, identification and biological characteristics of rat bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND: How to effectively harvest bone marrow mesenchymal stem cells with homogeneity is crucial for cartilage tissue engineering research. OBJECTIVE: To isolate and culture rat bone marrow mesenchymal stem cells and to observe biological characteristics of the cells. METHODS: Bone marrow mesenchymal stem cells were isolated from the femur and tibia of 30 Sprague-Dawley rats using the whole bone marrow adhesion method. Cell surface markers were identified using flow cytometry, cell proliferation ability was observed through cell counting, and cell survival rate was determined by MTT assay. RESULTS AND CONCLUSION: The harvested cells were mostly round and oval, which partially showed a triangular shape. Cell fusion appeared after 12-15 days of inoculation. The proliferation of passaged cells was accelerated. Two hours after passage, the cells evenly covered the bottom of the culture dish. With the increasing of cell generations, the cell proliferation ability was gradually decreased. The survival rates of cells at passages 1-5 were over 95%, significantly higher than those at passages 6 and 7 (P < 0.05). The harvested cells were negative for CD34, but positive for CD44. To conclude, the whole bone marrow adhesion method is effective to isolate rat bone marrow mesenchymal stem cells with stable growth and strong proliferation ability. Passage 5 cells can be selected as seed cells for cartilage tissue engineering.