骨髓间质干细胞体外增殖及诱导分化成肌样细胞实验研究

In vitro proliferation and differentiation into muscle-like cells of bone marrow mesenchymal stem cell

目的比较SD大鼠、Wistar大鼠和C57BL/6J小鼠骨髓间质干细胞在体外的增殖及在体外将骨髓间质干细胞诱导分化成肌样细胞的情况。方法体外分离成年SD大鼠、Wistar大鼠和C57BL/6J小鼠的骨髓干细胞,培养增殖,用5-氮杂胞苷诱导分化,使其向肌细胞定向分化。结果SD大鼠和Wistar大鼠的原代骨髓间质干细胞约10 d可达90%以上融合,平均3 d左右传代,C57BL/6J小鼠的骨髓间质干细胞在1周内可迅速增殖,1周后基本处于停滞状态,且细胞集落很少,难传代。SD大鼠的骨髓间质干细胞在含5-氮杂胞苷、两性霉素B和5%马血清的分化培养基作用下,可以分化为肌肉特异性抗原desmin和-αsarcomeric actin染色阳性反应的肌样细胞。结论不同种系的骨髓干细胞在体外用相同的培养基培养,可以具有截然不同的增殖能力;5-氮杂胞苷可以促使骨髓干细胞定向分化为肌样细胞。

Objective To compare in vitro proliferation abihty of bone marrow mesenchymal stem cells derived from SD rat, Wistar rat and C57BL/6J meuse and to differentiate bone marrow mesenchymal stem cells derived from SD rat into musclelike cells, Methods Bone marrow stem cells derived from SD rat, Wistar rat and C57BL/6J mouse were isolated and cultured. Bone marrow mesenchymal stem cells derived from SD rat were induced to differentiate into muscle-like cells by 5-azacytidine. Results Bone marrow mesenchymal stem cells derived from SD rat and Wistar rat reached 90% confluence in about 10 days and could be passaged in 3 days; Bone marrow mesenchymal stem cells derived from C57BL/6J mouse proliferate rapidly in 1 week but was prone to rest later that. The cell clone was very scares, It was very difficult to be passaged under the same culture conditions as SD and Wistar rat. The bone marrow mesenchymal stem cells derived from SD rat could be differentiated into muscle-like cells which expressed desmin and α-sarcomeric actin by the use of differentiation medium containing 5-azacytidine, amphotericin B and 5 % horse serum. Conclusion Bone marrow stem cells from different species show different proliferation abihty, 5-azacytidine can induce bone marrow mesenchymal stem cells to differentiate into muscle-like cells.