Functional isolation,culture and cryopreservation of adult human primary cardiomyocytes

作     者：Bingying Zhou Xun Shi Xiaoli Tang Quanyi Zhao Le Wang Fang Yao Yongfeng Hou Xianqiang Wang Wei Feng Liqing Wang Xiaogang Sun Li Wang Shengshou Hu 

作者机构：State Key Laboratory of Cardiovascular DiseaseFuwai HospitalNational Center for Cardiovascular DiseasesChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina Shenzhen Key Laboratory of Cardiovascular DiseaseFuwai Hospital Chinese Academy of Medical SciencesShenzhenShenzhenChina Department of Cardiac SurgeryFuwai HospitalNational Center for Cardiovascular DiseasesChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingChina Present address:18 Jinma Industrial ParkFangshan DistrictBeijingChina 

出 版 物：《Signal Transduction and Targeted Therapy》 (信号转导与靶向治疗（英文）)

年 卷 期：2022年第7卷第8期

页      面：3093-3108页

核心收录：

学科分类：0710[理学-生物学] 1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

基　　金：supported by the CAMS Initiative for Innovative Medicine Program(grants 2021-1-I2M-006 and 2017-I2M-1-003) the National Natural Science Foundation of China(grants 82070287 and 81700337) 

主　　题：cardiomyocytes unstable culture 

摘      要：Cardiovascular diseases are the most common cause of death *** modeling cardiac homeostasis,dysfunction,and drug response lies at the heart of cardiac *** human primary cardiomyocytes(hPCMs)are a promising cellular model,but unstable isolation efficiency and quality,rapid cell death in culture,and unknown response to cryopreservation prevent them from becoming a reliable and flexible in vitro cardiac *** the use of a reversible inhibitor of myosinⅡATPase,(-)-blebbistatin(Bleb),and multiple optimization steps of the isolation procedure,we achieved a 2.74-fold increase in cell viability over traditional methods,accompanied by better cellular morphology,minimally perturbed gene expression,intact electrophysiology,and normal neurohormonal *** optimization of culture conditions established a method that was capable of maintaining optimal cell viability,morphology,and mitochondrial respiration for at least 7 *** importantly,we successfully cryopreserved hPCMs,which were structurally,molecularly,and functionally intact after undergoing the freeze-thaw *** demonstrated greater sensitivity towards a set of cardiotoxic drugs,compared to human-induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs).Further dissection of cardiomyocyte drug response at both the population and single-cell transcriptomic level revealed that hPCM responses were more pronouncedly enriched in cardiac function,whereas hiPSC-CMs responses reflected cardiac ***,we established a full set of methodologies for the efficient isolation and prolonged maintenance of functional primary adult human cardiomyocytes in vitro,unlocking their potential as a cellular model for cardiovascular research,drug discovery,and safety pharmacology.