中国实验动物学报2024，Vol.32Issue(4)：435-443,9.DOI:10.3969/j.issn.1005-4847.2024.04.003
成年小鼠心肌细胞腺病毒转染及缺氧/复氧诱导损伤模型的建立
Models of adenoviral transfection and hypoxia/reoxygenation-induced injury in AMCMs of adult mouse cardiomyocytes
摘要
Abstract
Objective To construct models of viral transfection and hypoxia/reoxygenation induced cellular injury in adult mouse cardiomyocytes(AMCMs)isolated using a non-Langendorff method.Methods AMCMs were isolated,extracted,sedimented,and plated using a non-Langendorff method.The morphology and survival rate of the isolated cells were evaluated 2,24,48 and 72 h after plating,and their integrity was observed by immunofluorescence staining for α-actinin.The isolated AMCMs were infected with adenoviruses carrying an RFP-expressing vector and fluorescence images were obtained at 36 and 48 h post-infection and used to calculate transfection efficiency.The cells were cultured under hypoxic conditions for 45 min,reoxygenated for 24 h,and then stained with propidium iodide(PI)to verify establishment of the hypoxia/reoxygenation injury model.Results The survival rates of AMCMs at 2,24 and 48 h after plating were comparable,but survival was significantly reduced at 72 h.The integrity of the AMCMs was good and＞80%of the cells were transfected with adenovirus at 48 h.After hypoxia/reoxygenation treatment,42%of cells were stained by PI,suggesting successful establishment of the AMCM injury model.Conclusions In this study,we developed a non-Langendorff method for the fast and easy isolation of AMCMs with high cell viability.The isolated cells can be efficiently infected with adenovirus and respond to hypoxia/reoxygenation injury.These findings provide a systematic method for isolating AMCMs and for applying gene modification and hypoxia/reoxygenation injury in these cells.关键词
AMCMs提取/腺病毒转染/缺氧/复氧Key words
AMCMs extraction/adenovirus transfection/hypoxia-reoxygenation分类
生物科学引用本文复制引用
李笑汝,殷倩,姚新叶,刘佳,张小玉,张依曼,来宝长,马强,王一东,田红燕..成年小鼠心肌细胞腺病毒转染及缺氧/复氧诱导损伤模型的建立[J].中国实验动物学报,2024,32(4):435-443,9.基金项目
国家自然科学基金青年基金(81900256),西安交通大学第一附属医院院基金(2018QN-04). Funded by National Natural Science Foundation of China(81900256),Research and Development Fund of the First Affiliated Hospital of Xi'an Jiaotong University(2018QN-04). （81900256）
