摘要
目的 建立成年大鼠心脏成纤维细胞的离体培养模型。方法 以组织块消化法分离细胞,以差数贴壁法去除其它细胞,常规培养、传代,免疫组化染色进行细胞鉴定并绘制生长曲线观察细胞传代后成纤维细胞的基本特征及生长特性的变化。结果 第2～4代细胞生长良好、稳定,波形蛋白染色阳性,Ⅷ因子相关抗原及肌动蛋白染色阴性。结论 组织块消化法是一种适用于成年大鼠心脏成纤维细胞分离培养的简便方法,第2～4代细胞适于实验。
Objective To establish a model of culture cardiac fibroblasts(CFs) from Wistar adult rat ventricule of in vitro and observe the growth properties of CFs in vitro. Methods Ventricular tissue was minced into small pieces and then subjected to digestion. Dispersed cells were plated onto tissue culture dishes and incubated for 0.5 h at 37℃ under 5% CO2. At the end of this period, the unattached cells were discarded, and attached cells(CFs) were cultured coutinaously. The fibroblastic nature of cell lines was determined by immunohistochemical method and cell growth curve was plotted. Results Immunocytochemical staining showed that the cells in culture at passage two-passage four were stained positive for vimentin and negative for muscle actin and factor Ⅷ. And the cells in culture at passage two-passage four also displayed stable growth feature.Conclusion This technique is suitable for culturing cardiac flbroblasts of adult rat and the 2nd-4th passage of culture is suitable for experimental work.
