title: Chemical-defined and albumin-free generation of human atrial and ventricular myocytes from human pluripotent stem cells.
abstract: Most existing culture media for cardiac differentiation of human pluripotent stem cells (hPSCs) contain significant amounts of albumin. For clinical transplantation applications of hPSC-derived cardiomyocytes (hPSC-CMs), culturing cells in an albumin containing environment raises the concern of pathogen contamination and immunogenicity to the recipient patients. In addition, batch-to-batch variation of albumin may cause the inconsistent of hPSC cardiac differentiation. Here, we demonstrated that antioxidants l-ascorbic acid, trolox, N-acetyl-l-cysteine (NAC) and sodium pyruvate could functionally substitute albumin in the culture medium, and formulated an albumin-free, chemical-defined medium (S12 medium). We showed that S12 medium could support efficient hPSC cardiac differentiation with significantly improved reproducibility, and maintained long-term culture of hPSC-CMs. Furthermore, under chemical-defined and albumin-free conditions, human-induced pluripotent stem cells (hiPSCs) were established, and differentiated into highly homogenous atrial and ventricular myocytes in a scalable fashion with normal electrophysiological properties. Finally, we characterized the activity of three typical cardiac ion channels of those cells, and demonstrated that hPSC-derived ventricular cardiomyocytes (hPSC-vCMs) were suitable for drug cardiac safety evaluation. In summary, this simplified, chemical-defined and albumin-free culture medium supports efficient generation and maintaining of hPSC-CMs and facilitates both research and clinical applications of these cells.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Pei
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Jiang
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Shuyun
authors.initials: S
authors.lastname: Bai
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Henghua
authors.initials: H
authors.lastname: Cao
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Luyang
authors.initials: L
authors.lastname: Tian
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Biology Unit, Research Division, WuXi AppTec Co., Ltd., Shanghai 200131, China.
authors.forename: Chuanxiu
authors.initials: C
authors.lastname: Yang
authors.affiliation: Biology Unit, Research Division, WuXi AppTec Co., Ltd., Shanghai 200131, China.
authors.forename: Haiheng
authors.initials: H
authors.lastname: Dong
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China; Medical School of University of Chinese Academy of Sciences, Beijing 100101, China.. Electronic address: yuema@ibp.ac.cn.
authors.forename: Yue
authors.initials: Y
authors.lastname: Ma
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000975:Antioxidants / Q000494:pharmacology; D051837:COUP Transcription Factor II / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D009543:Nifedipine / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010880:Piperidines / Q000494:pharmacology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D000975:Antioxidants; D051837:COUP Transcription Factor II; D003470:Culture Media; D018398:Homeodomain Proteins; D009543:Nifedipine; D010880:Piperidines; D011725:Pyridines; D014212:Tretinoin; C485533:IRX4 protein, human; C063968:E 4031; C095569:NR2F2 protein, human
keywords: Albumin; Antioxidants; Cardiac subtypes specification; Drugs; Human pluripotent stem cells (hPSCs)