Functional isolation, culture and cryopreservation of adult human primary cardiomyocytes

• DOI: 10.1038/s41392-022-01044-5
• Publication Date: 2022-07-29T14:45:49Z
• Authors: 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
• Keywords: Viability assay

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