摘要
目的培养成纤维细胞,建立不同种属来源的胸主动脉成纤维细胞的培养方法并比较其形态差异。方法用消化法和贴壁法分离不同来源的成纤维细胞,利用差速培养法纯化成纤维细胞,并对所培养的细胞进行倒置显微镜观察和苏木素-伊红染色,观察成纤维细胞形态,并对培养细胞行波形蛋白免疫染色鉴定。结果接种24h后细胞全部贴壁,48h后人胸主动脉成纤维细胞体积增大并伸出伪足,形状以梭形或不规则形为主;而大鼠胸主动脉的成纤维细胞生长的相对较慢。结论用Ⅰ型胶原消化动脉的成纤维细胞效果较好,用差速生长纯化的成纤维细胞可用于实验研究。
Objective To set up the method for the culture of fibroblasts from different sources. Methods Human fibroblast cells were isolated through enzyme digestion and adherence, and then purified by differential attachmcnt technique. The morphology and phenotypc characterization of cultured fibroblasts was inspected by light microscopy after HE and immunocytochemical staining for vimentin. Results 24 h after inoculation, all the cells had attached. 48 h after inoculation, the volume of human artery fibroblast cell enlarged and pseudopodia stretched on, mostly in the shape of irregular or clostridial form. Rat thoracic aorta fibroblast cells grow relatively slower. Conclusion In the primary culture of human and rat thoracic aortic fibroblast cells, it is recommended to digest by type- I collagen and purify by differential attachment technique.
出处
《海南医学》
CAS
2012年第6期23-25,共3页
Hainan Medical Journal
关键词
胸主动脉
成纤维细胞
原代培养
Thoracic aorta
Fibroblast cell
Primary culture
