Journal
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY
Volume 85, Issue -, Pages 79-88Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.yjmcc.2015.05.003
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Funding
- United States National Institutes of Health [HL88635, HL71670, HL108173, NIH HL104040, U01HL099773]
- American Heart Association [13IRG13680003, 12SDG12050597, 12POST12080080]
- Veterans Administration Merit Grant [BX000771]
- National Center for Research Resources Grant [UL1 RR024975]
- National Center for Advancing Translational Sciences Grant [UL1 TR000445]
- NIH [CA68485, DK20593, DK58404, DK59637, EY08126]
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Cardiomyocytes (CMs) derived from human induced pluripotent stem cells (hiPSCs) are being increasingly used to model human heart diseases. hiPSC-CMs generated by earlier aggregation-based methods (i.e., embryoid body) often lack functional sarcoplasmic reticulum (SR) Ca stores characteristic of mature mammalian CMs. Newer monolayer-based cardiac differentiation methods (i.e., Matrigel sandwich or small molecule-based differentiation) produce hiPSC-CMs of high purity and yield, but their Ca handling has not been comprehensively investigated. Here, we studied Ca handling and cytosolic Ca buffering properties of hiPSC-CMs generated independently from multiple hiPSC lines at Stanford University, Vanderbilt University and University of Wisconsin-Madison. hiPSC-CMs were cryopreserved at each university. Frozen aliquots were shipped, recovered from cryopreservation, plated at low density and compared 3-5 days after plating with acutely-isolated adult rabbit and mouse ventricular CMs. Although hiPSC-CM cell volume was significantly smaller, cell capacitance to cell volume ratio and cytoplasmic Ca buffering were not different from rabbit-CMs. hiPSC-CMs from all three laboratories exhibited robust L-type Ca currents, twitch Ca transients and caffeine-releasable SR Ca stores comparable to adult CMs. Ca transport by sarcoendoplasmic reticulum Ca ATPase (SERCA) and Na/Ca exchanger (NIX) was similar in all hiPSC-CM lines, but slower compared to rabbit-CMs. However, the relative contribution of SERCA and NCX to Ca transport of hiPSC-CMs was comparable to rabbit-CMs. Ca handling maturity of hiPSC-CMs increased from 15 to 21 days post-induction. We conclude that hiPSC-CMs generated independently from multiple iPSC lines using monolayer-based methods can be reproducibly recovered from cryopreservation and exhibit comparable and functional SR Ca handling. (C) 2015 The Authors. Published by Elsevier Ltd.
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