采用组织块法培养心房颤动患者心房成纤维细胞  

A model establishment for cell culture of human myocardial fibroblasts with tissue culture in atrial fibrillation patients

作　　者：张建成[1] 程显禄[1] 黄宇坚[1] 许春萱[1] 

机构地区：[1]福建医科大学省立临床医学院心血管内科,福建福州350004

出　　处：《中国心脏起搏与心电生理杂志》2009年第6期506-508,共3页Chinese Journal of Cardiac Pacing and Electrophysiology

基　　金：福建省卫生厅重点创新项目(项目编号:2007-CX-1)

摘　　要：目的建立心房颤动（简称房颤）患者心房肌成纤维细胞培养模型。方法通过心脏外科手术获得房颤患者右心耳组织30mg，对经典的大鼠组织块培养法作改进，采用组织块放入75％酒精中浸泡1min，培养基配方和植块方法改进，第二代传代时采用差速贴壁法除去内皮细胞，培养房颤患者心房肌成纤维细胞。结果形态学观察显示，心肌成纤维细胞生长较慢，3天左右开始从组织块边缘爬出，10天呈接合状态，细胞呈梭形，细胞排列较紧密，有的交叉重叠生长，胞核较大，呈椭圆形，通常含1～3个核，胞浆透明，胞体较大，无搏动；免疫细胞化学染色可见，心肌成纤维细胞波形蛋白呈强阳性反应，第八因子呈阴性反应；成功培养的心肌成纤维细胞纯度达95％以上。结论采用组织块法可成功培养人心房的心肌成纤维细胞，且简单、易行。Objective To establish the model for cell culture of human myocardial fibroblasts in atrial fibrillation patients. Methods humann myocardial fibroblasts were isolated and cultured with a improved technique of tissue culture. Tissue was immersed into 75% alcohol 1 minute, improved explant medium formulations and planting ways; The second Cell Subculture used differential adhesion method to remove endothelial cells. Results Morphology of the cells were observed using the inverted microscope. Myocardial fibroblasts growed slowly, abaut three days climbing from the edge of the tissue and ten days showing fusion state, closely arranging, and some overlaping; It was spindle-shaped, larger, transparent in the cytoplasm, and it was much larger nucleus that was oval-shaped, usually with 1 or 3 nucleus and non-spontaneous beat. Identification of immunocytochemical staining showed that cardiac fibroblasts showed strongly positive reaction to vimentin and negative to factor VIII,which showed the purity of cells cultured was more than percent 95. Conclusion The model of cell culture of human myocardial fibroblasts can be successfully established and the method is simple and easy.

关 键 词：心血管病学 心房颤动 心肌成纤维细胞 细胞培养 

分 类 号：R541.75[医药卫生—心血管疾病]