title: Comparing mouse and human pluripotent stem cell derived cardiac cells: Both systems have advantages for pharmacological and toxicological screening.
abstract: Pluripotent stem cells offer an unparalleled opportunity to investigate cardiac physiology, pharmacology, toxicology and pathophysiology. In this paper we describe the use of both mouse (Nkx2-5(eGFP/w)) and human (NKX2-5(eGFP/w)) pluripotent stem cell reporter lines, differentiated toward cardiac lineage, for live single cell high acquisition rate calcium imaging. We also assess the potential of NKX2-5(eGFP/w) cardiac lineage cells for use toxicological screening as well as establish their sensitivity to a shift between low and high oxygen environments. Differentiated mouse Nkx2-5(eGFP/w) cells demonstrated a wide range of spontaneous oscillation rates that could be reduced by ryanodine (10μM), thapsigargin (1μM) and ZD7288 (10μM). In contrast human NKX2-5(eGFP/w) cell activity was only reduced by thapsigargin (1μM). Human cell survival was sensitive to the addition of trastuzumab and doxorubicin, while the switch from a low to a high oxygen environment affected oscillation frequency. We suggest that the human NKX2-5(eGFP/w) cells are less suitable for studies of compounds affecting cardiac pacemaker activity than mouse Nkx2-5(eGFP/w) cells, but are very suitable for cardiac toxicity studies.
authors.affiliation: Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), Melbourne, Australia.
authors.forename: E L
authors.initials: EL
authors.lastname: Lagerqvist
authors.affiliation: Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), Melbourne, Australia.
authors.forename: B A
authors.initials: BA
authors.lastname: Finnin
authors.affiliation: Murdoch Children's Research Institute, Monash University, Clayton, Victoria, Australia.
authors.forename: D A
authors.initials: DA
authors.lastname: Elliott
authors.affiliation: Murdoch Children's Research Institute, Monash University, Clayton, Victoria, Australia.
authors.forename: D J
authors.initials: DJ
authors.lastname: Anderson
authors.affiliation: Cardiovascular Research Center, Massachusetts General Hospital, Boston, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Wu
authors.affiliation: Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), Melbourne, Australia.
authors.forename: C W
authors.initials: CW
authors.lastname: Pouton
authors.affiliation: Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University (Parkville Campus), Melbourne, Australia. Electronic address: john.haynes@monash.edu.
authors.forename: J M
authors.initials: JM
authors.lastname: Haynes
mesh_terms: D000818:Animals; D001683:Biological Clocks / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D051379:Mice; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*
chemical_list: D010100:Oxygen
keywords: Calcium imaging; Ischemia; Oscillation frequency; Stem cell-derived cardiomyocytes; Toxicity