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目的:将体外培养并纯化的自体骨骼肌成肌细胞植入心肌梗死区,观察其心脏功能有无改善及改善程度,以探讨成肌细胞移植治疗的机制。方法:采用反复差速贴壁法体外培养、纯化及鉴定新西兰兔自体成肌细胞。结扎冠状动脉致局部心肌梗死后2周,分别将成肌细胞和对照的细胞培养液注入心肌梗死区,4周后观察心功能改善情况,鉴定心肌梗死边缘区毛细血管密度及植入细胞的形态。结果:体外培养获得高纯度(85.4%±6.7%)Desm in阳性的自体骨骼肌成肌细胞。经Buxco系统有创心功能测定显示自体成肌细胞移植组较对照组的左心室等容收缩期室内压最大上升速率[+dp/dtmax,(1 217.77±89.91)mmHg/s vs.(897.83±70.04)mmHg/s]和左心室等容舒张期室内压最大下降速率[-dp/dtmax,(-1 174.58±91.5)mmHg/s vs.(-753.67±69.66)mmHg/s]均有所改善。免疫组织化学和电镜证实植入的细胞为肌源性且细胞移植区域的毛细血管密度较对照组高。结论:采用反复差速贴壁法培养可在体外获得高纯度骨骼肌成肌细胞,植入成肌细胞能增加心肌收缩力,改善心功能并促进心肌梗死区域的毛细血管数量增加。
OBJECTIVE: To implant autologous skeletal muscle myoblasts cultured in vitro and purified into myocardial infarction area to observe the improvement of cardiac function and the degree of improvement, so as to explore the mechanism of myoblast transplantation. Methods: In vitro culture, purification and identification of autologous myoblasts from New Zealand rabbits were performed by repeated differential adherence method. Two weeks after the local myocardial infarction, the myoblasts and the control cells were infused into the myocardial infarction area. The improvement of cardiac function was observed after 4 weeks, and the capillary density and morphology of the implanted cells in the marginal zone of myocardial infarction were identified. RESULTS: Desm in-positive autologous skeletal muscle myoblasts were obtained in vitro with high purity (85.4% ± 6.7%). The maximal rate of increase of left ventricular isovolumic systolic pressure in autologous myoblast transplantation group was higher than that in control group [+ dp / dtmax (1 217.77 ± 89.91) mmHg / s vs. (897.83 ± 70.04) mmHg / s], and the maximal rate of decline in the left ventricular isovolumic diastolic pressure [-dp / dtmax, (-1 174.58 ± 91.5) mmHg / s versus 753.67 ± 69.66 mmHg / s] . Immunohistochemistry and electron microscopy confirmed that the implanted cells were myogenic and the capillary density in the cell-grafted area was higher than that in the control group. CONCLUSION: High purity skeletal myoblasts can be obtained in vitro by repeated differential adherence method. Implantation of myoblasts can increase myocardial contractility, improve cardiac function and increase the number of capillaries in myocardial infarction area.