冠心病病人骨髓间充质干细胞的生物学特征评价

Biological characteristics of bone marrow derived mesenchymal stem cells from patients with coronary artery disease

目的 确立胸骨骨髓间充质干细胞分离和培养的方法 ,评价其生物学特征。方法 骨髓取自行冠状动脉旁路移植术的冠心病病人胸骨切口 ,用Percoll液分离骨髓MSCs,体外培养扩增 ,观察细胞生长特性 ,流式细胞仪检测骨髓MSCs表型特征、细胞周期。结果 贴壁的骨髓MSCs多数呈梭形。骨髓MSCs能连续传 15代以上 ,但是第 5代以后或老年病人的细胞增殖速度减慢。骨髓MSCs的表型特征为CD2 9、CD4 4阳性 ,CD34、CD4 5阴性。多数骨髓MSCs在细胞周期的G0 /G1期。结论 冠心病病人手术时从胸骨切口取骨髓并分离培养骨髓MSCs的方法可行 ,骨髓MSCs具有较好的增殖更新潜能 ,是细胞心肌成形术中理想的细胞来源。

Objective To establish a method for the isolation and culture of bone marrow derived mesenchymal stem cells (MSCs) from patients with coronary artery disease and investigated its biological characteristics. Methods Bone marrow was extracted from patients' sternum when they were operated for coronary artery bypass graft. MSCs were separated from bone marrow by density gradient centrifugation in Percoll solution and seeded in culture medium supplemented with 10% fetal bovine serum. The morphology and growth characteristic of MSCs were observed in primary and passage generation culture with inverted microscope. The cell phenotype and cell cycle of MSCs were analyzed by flow cytometry. The growth curves were compared between different passages and ages. Results Most adherent MSCs showed spindle morphology whatever passages they were. The MSCs could be expanded for 15 passages. The proliferative potential of MSCs over the fifth passage or from old patients decreased gradually. The MSCs in primary and passage were positive for CD29 and CD44, but negative for CD34 and CD45. Analysis for cell cycle showed most MSCs in G_0/G_1 phase. Conclusion The method of isolation and culture of sternum bone marrow derived MSCs from patients with coronary artery disease is feasible, the MSCs have self-renewal capacity and might be ideal for cellular cardiomyoplasty.