骨髓间充质干细胞体外扩增和向心肌细胞分化的实验研究

In vitro proliferation and cardiomyocytes induction of bone marrow mesenchymal stem cells

①目的 探讨兔骨髓间充质干细胞的体外分离纯化、培养扩增和向心肌细胞诱导分化的条件。②方法 取兔股骨骨髓 ,利用密度为 1 .0 73g/mL的Percoll分离骨髓细胞 ,以含 1 0 %胎牛血清的低糖DMEM培养基培养与扩增骨髓间质干细胞。取第二代和第三代的骨髓间质干细胞 ,以含有不同浓度的 5 -氮杂胞苷诱导培养基向心肌细胞诱导分化。③结果 诱导后的细胞呈现典型的心肌细胞样改变 ,免疫组化染色显示其α -横纹肌动蛋白阳性。④结论 骨髓间质干细胞在体外具有向心肌细胞分化的潜能。

Objective This study investigated the culture and in vitro proliferation of mesenchymal stem cells as well as requirements for cardiomyocytes differentiation of mesenchymal stem cells Methods Bone marrow was aspirated from the rabbit tibia. Mesenchymal stem cells in each bone marrow samples were obtained by gradient centrifugation with Percoll at a density of 1.073g/ml and then cultured and expanded with lower glucose Dulbecco's modified Eagle's medium (L-DMEM) containing 10% fetal bovine serum (FBS). Mesenchymal stem cells derived from the second and the third passages were cultured for 14 and 21 days in the media containing 5-azaytidine of various combinations. Results The induced mesenchymal stem cells changed their morphology to that of cardiomyocyte type. Immunohistochemical studies proved the cells Alpha-Sarcomeric Actin expression. Conclusion Mesenchymal stem cells were induced to differentiate along cardiomyocytes lineage.