摘要
目的 建立风心病心肌细胞体外培养方法 ,观察其胶原合成变化 .方法 采用快速贴壁法对正常成人 7例和风心患者 15例左室乳头肌心肌细胞进行原代培养 ,观察培养期间其形态、超微结构、活性及胶原合成的变化 .结果 培养的成人心肌细胞短而粗呈杆状 ,无明显自主收缩 ;48h后杆状细胞率为 ( 87.2± 4.1) % ,细胞存活率为 ( 92 .8± 4.1) % ,两者随培养时间延长而降低 ;透射电镜显示杆状细胞超微结构正常 ;免疫组化染色显示风心病心肌细胞 , 型胶原呈阳性表达 ,正常组呈阴性或弱阳性表达 ;胶原合成测定结果显示风心组[2 ,3- 3H]-脯胺酸摄取量显著高于正常组 .结论 我们建立的成熟心肌细胞分离和培养方法可靠 ,培养心肌细胞保持了在体的杆状形态 ,风心病心肌细胞培养 48h后胶原合成表型仍存在 。
AIM To establish a method for culturing adult cardiomyocytes with rheumatic heart disease (ACMR) in vitro, and to study the changes in its collagen synthesis. METHODS The adult cardiomyocytes (ACM) of left ventricular papillary muscle of seven normal persons and fifteen patients with rheumatic heart disease were cultured primarily in vitro by means of rapid attachment. The changes in morphology, ultrastructure, viability and collagen synthesis of ACM were studied in culture. RESULTS The cultured ACM were rodshaped without spontaneous contractility. The ratio of rodshaped cells and viable cells was (87.2± 4.1 )% and (92.8±4.1)% after 48 h in culture, and the ultrastructure of rodshaped cells was similar to that of the cardiac tissue in vivo. Immunohistochemistry staining showed that the expression of type Ⅰ, Ⅲ collagen in cultured ACMR was positive, while that of the control group was negative or weakly positive. The collagen synthesis of ACMR cultured in medium with normal human or RHD patient's serum was significantly higher than that of the control group. CONCLUSION The method for isolating and culturing ACM is reliable, and the phenotype of collagen synthesis of ACMR still existed after 48 h in culture. This model provides an effective experimental method for studying the mechanism of myocardial fibrosis in rheumatic heart disease that cardiomyocytes participate in.
出处
《第四军医大学学报》
2000年第5期543-546,共4页
Journal of the Fourth Military Medical University
基金
国家自然科学基金资助项目!(39870790)
关键词
风湿性心脏病
成熟心肌细胞
体外培养
胶原
rheumatic heart disease
adult cardiomyocytes
culture in vitro
collagen
