SD乳鼠原代心脏成纤维细胞培养及鉴定  

Culture and Identification of Primary Cardiac Fibroblasts in SD Neonatal Mice

作　　者：杨艳[1] 欧阳伟炜[1,2] 苏胜发[1,2] 马筑[1,2] 李青松[1,2] 耿一超 卢冰[1,2] YANG Yan;OUYANG Weiwei;SU Shengfa;MA Zhu;LI Qingsong;GENG Yichao;LU Bing(Department of Oncology,the Affiliated Hospital of Guizhou Medical University,Department of Oncology,Guizhou Provincial Cancer Hospital,Guiyang 550004,Guizhou,China;Department of Oncology,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学附院肿瘤科,贵州省肿瘤医院肿瘤科,贵州贵阳550004 [2]贵州医科大学肿瘤学教研室,贵州贵阳550004

出　　处：《贵州医科大学学报》2020年第4期427-431,共5页Journal of Guizhou Medical University

基　　金：贵州省科技厅课题[黔科合LH字(2014)7135];贵州省教育厅创新群体重大研究项目[黔教合KY字(2016)032];国家自然科学基金地区科学基金项目(81660507)。

摘　　要：目的:建立一种乳鼠心脏成纤维细胞分离、培养及鉴定的方法。方法:选取新出生的SD乳鼠,用胰蛋白酶和Ⅱ型胶原酶消化组织,差速贴壁法分离纯化心脏成纤维细胞;倒置显微镜观察细胞形态特征及生长情况,并进行Vimentin免疫组化对培养的原代细胞进行鉴定,观察原代细胞生长中出现污染情况。结果:差速贴壁60~90 min后镜下可见圆形细胞贴壁,2 d后可见细胞呈梭形、扁平形、三角形或多角形,且细胞数量增多、核分裂相增多;第3~4天时细胞生长约90%,免疫组化见细胞染色达90%以上,在原代细胞培养48 h时出现细菌污染。结论:成功建立一种稳定、有效的心脏成纤维细胞培养方法。Objective:To establish a method for the isolation,culture and identification of cardiac fibroblasts in SD neonatal mice.Methods:The SD newborn mice were selected and the tissues were digested with trypsin and type II collagenase,and the cardiac fibroblasts were isolated and purified by rapid adherence.Cardiac fibroblasts and myocardial cells were isolated with differential adhesion method.The morphological changes and grows of cardiac fibroblasts were observed under inverted microscope.The primary cells are identified by Vimentin immunochemistry staining and the contamination in primary cell growth was observed.Results:After 60~90 minutes of differential adhesion,adherent fibroblasts formed spherical cell mass and cells were spindle-shaped and proliferated rapidly after 2 days.Cells were confluent after 3~4 days and nuclear fission increased.The positive rate of Vimentin immunochemistry staining were nearly up to 90%.Bacterial contamination occurred during primary cell culture at 48 h.Conclusion:This method is economical and stable to isolate cardiac fibroblasts with high activity and high purity from adult mice.

关 键 词：心脏 成纤维细胞 形态发生 免疫组织化学 原代培养 鉴定 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] Q954.6[医药卫生—基础医学]