摘要
目的建立稳定的成年大鼠心肌细胞分离和培养方法,以便进行成年大鼠心肌细胞收缩功能的研究.方法将成年大鼠心脏挂于Langendorff装置上灌流,胶原酶消化分离成年大鼠心肌,无血清悬浮培养心肌细胞.光镜观察,结合形态学和收缩功能评价心肌细胞.结果新分离的心肌细胞的收获量为(4～6)×106个,杆状和圆形,其中长杆状细胞大于75%,横纹清晰,收缩幅度(12.00±2.68)%.无血清培养24h后,细胞保持完整的形态结构,长杆状心肌细胞占总数的70%以上,收缩幅度(11.78±1.59)%,与刚培养时的细胞相比,无显著差别(P＞0.05).结论通过本方法可以对成年大鼠心肌细胞进行良好的分离培养.
Objective To build a stable method for isolation and culture of adult rat cardiomyocytes. Methods The isolated adult rat heart was mounted on to the Langendorff apparatue for perfusion and hallogenase digestion. The treated heart was processed to get the suspension of single cardiomyocytes for culture in serum - free DMEM medium. The morphology and contractile function of the cardiomyocytes were studied under optical microscope to compare the fresh and cultured cardiomyocytes. Results The total of freshly isolated adult rat cardiomyocytes was （4 - 6） × 10^6 and more than 75 % of them were rod - shaped cells with clear cross - striations. The amplitude of unloaded shortening in fresh cardiomyocytes was （ 12.00 ± 2.68） % . In the 24 h cardiomyocyte culture, the rod - shaped cells accounted for more than 70% and their amplitude of unloaded shortening was （ 11.78 ± 1.59） %. No differences were noticed between the freshly isolated cells and the cultured ones （ P 〉 0.05 ）. Conclusion Adult rat ventricular cardiomycytes can be well isolated and cultured by using the stated method.
出处
《徐州医学院学报》
CAS
2005年第5期393-396,共4页
Acta Academiae Medicinae Xuzhou
基金
江苏省教育厅自然科学基金资助项目(02KJB0008)
