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Publication
Comparable calcium handling of human iPSC-derived cardiomyocytes generated by
multiple laboratories.
Authors Hwang HS, Kryshtal DO, Feaster TK, Sánchez-Freire V, Zhang J, Kamp TJ, Hong CC,
Wu JC, Knollmann BC
Submitted By Submitted Externally on 11/3/2015
Status Published
Journal Journal of molecular and cellular cardiology
Year 2015
Date Published 8/1/2015
Volume : Pages 85 : 79 - 88
PubMed Reference 25982839
Abstract 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 3-5days 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 (NCX) 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
21days 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.




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