| 摘要: |
| 目的:体外分选纯化人骨髓多潜能成体祖细胞(MAPCs),诱导分化,观察其分化后细胞移植治疗心肌梗死的疗效,初步探讨MAPCs移植治疗心肌梗死的可行性.方法:取适量志愿者骨髓采用梯度密度离心法分离获取单个核细胞并行贴壁培养,将获取的细胞通过CD45、GlyA免疫微磁珠负分选(MACS)分离出MAPCs;锥虫蓝拒染试验测定分选后细胞活力,流式细胞仪鉴定分选后细胞纯度;传代后的细胞与10 μmol·L-1浓度5-氮杂胞苷(5-aza)共孵育24 h,通过透射电镜对诱导后的细胞进行鉴定,并将诱导后的MAPCs移植于兔心肌梗死部位,观察其对兔心功能的改善情况. 结果:MACS分选前后MAPCs细胞活力分别为(96.7±1.7)﹪、(96.0±2.4)﹪,无显著差异;获取的CD45-、GlyA-细胞纯度大于98﹪.分选后传代的MAPCs经5-aza诱导后,透射电镜检查可见大量的粗大肌丝;细胞移植治疗心肌梗死兔4周后,MAPCs移植组左室射血分数、左室侧壁运动幅度、左室收缩期室壁增厚率,以及左心室最大压力上升速率 ( dp/dt),均明显高于心梗模型组和空白对照组(P＜0.05或0.01),但低于假手术组(P＜0.05);空白对照组与心梗模型组相比无显著差异.结论:MACS分离出的高纯度人MAPCs在体外经5-aza诱导后能够向肌源性细胞分化,兔心梗区内植入诱导后的MAPCs能够改善心功能. |
| 关键词: 人骨髓多潜能成体祖细胞、细胞分化、细胞移植、心肌梗塞 |
| DOI：10.3724/SP.J.1008.2006.00853 |
| 投稿时间：2006-06-19修订日期：2006-07-20 |
| 基金项目: |
|
| Differentiated bone marrow-derived multipotent adult progenitor cells in treatment of rabbit cardiac infarction |
| WANG Xiao-hua,WU Zhong-hai,TANG Li-jun |
| (南方医科大学基础医学部生理学教研室,广州,510515;解放军福州总医院普通外科,福州,350002) |
| Abstract: |
| Objective：To study the biological characteristics and the differentiation ability of multipotent adult progenitor cells （MAPCs） in vitro and to observe the clinical outcome of MAPC differentiated cells in treatment of ischemic heart disease （IHD） in rabbits, so as to assess the feasibility of transplanting MAPCs in treatment of cardiac infarction. Methods： The bone marrow mononuclear cells were separated from volunteer bone marrow by gradient density centrifugation. The mononuclear cells were separated in CD45 and GlyA fractions by magnetic cell sorting （MACS）. The viability of MAPCs was assessed by trypan-blue assay and their purity were detected by flow cytometry. After cultured with 5-azacytidine （10 μmol/L） for 24 h, MAPCs were subjected to transmission electron microscopic observation. The myogenic cells differentiated from MAPCs were transplanted into the myocardial infarction sites in rabbits and the improvement of their cardiac function was evaluated. Results： The viability of unpurified MAPCs were （96.7±1.7）%, which was not significantly different from that of the purified MAPCs （96.0±2.4） %. The purity of MACS-sorted CD45^- and GlyA^- cells was 〉98%. Thick myofibers were observed by transmission electron microscopy after induction with 5-aza. Four weeks after transplantation, the left ventricle ejection fraction, the movement extent of left ventricle, left ventricle systolic wall thickening and dp/dt of rabbits were all higher than those of the control group and cardiac infarction model group （P〈0.05 or 0.01）, but lower than those of the sham operation group （P〈0.05）. Conclusion： The human MAPCs can differentiate into myogenic cells in the presence of 5-aza. The cardiac function of cardiac infarcted rabbits can be improved by MAPCs transplantation |
| Key words: multipotent adult progenitor cells from bone marrow cell differentiation cell transplantion myocardial infarction |
