TY - JOUR
T1 - Generation of ventricular-like hipsc-derived cardiomyocytes and high-quality cell preparations for calcium handling characterization
AU - Oh, Jae Gyun
AU - Dave, Jaydev
AU - Kho, Changwon
AU - Stillitano, Francesca
N1 - Funding Information:
This research was supported by AHA Scientist Development Grant 17SDG33700093 (F.S.); Mount Sinai KL2 Scholars Award for Clinical and Translational Research Career Development KL2TR001435 (F.S.); NIH R00 HL116645 and AHA 18TPA34170460 (C.K.).
Publisher Copyright:
© 2020 Journal of Visualized Experiments.
PY - 2019/1
Y1 - 2019/1
N2 - Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) provide a valuable human source for studying the basic science of calcium (Ca2+) handling and signaling pathways as well as high-throughput drug screening and toxicity assays. Herein, we provide a detailed description of the methodologies used to generate high-quality iPSC-CMs that can consistently reproduce molecular and functional characteristics across different cell lines. Additionally, a method is described to reliably assess their functional characterization through the evaluation of Ca2+ handling properties. Low oxygen (O2) conditions, lactate selection, and prolonged time in culture produce high-purity and high-quality ventricular-like cardiomyocytes. Similar to isolated adult rat cardiomyocytes (ARCMs), 3-month-old iPSC-CMs exhibit higher Ca2+ amplitude, faster rate of Ca2+ reuptake (decay-tau), and a positive lusitropic response to β-adrenergic stimulation compared to day 30 iPSC-CMs. The strategy is technically simple, cost-effective, and reproducible. It provides a robust platform to model cardiac disease and for the large-scale drug screening to target Ca2+ handling proteins.
AB - Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) provide a valuable human source for studying the basic science of calcium (Ca2+) handling and signaling pathways as well as high-throughput drug screening and toxicity assays. Herein, we provide a detailed description of the methodologies used to generate high-quality iPSC-CMs that can consistently reproduce molecular and functional characteristics across different cell lines. Additionally, a method is described to reliably assess their functional characterization through the evaluation of Ca2+ handling properties. Low oxygen (O2) conditions, lactate selection, and prolonged time in culture produce high-purity and high-quality ventricular-like cardiomyocytes. Similar to isolated adult rat cardiomyocytes (ARCMs), 3-month-old iPSC-CMs exhibit higher Ca2+ amplitude, faster rate of Ca2+ reuptake (decay-tau), and a positive lusitropic response to β-adrenergic stimulation compared to day 30 iPSC-CMs. The strategy is technically simple, cost-effective, and reproducible. It provides a robust platform to model cardiac disease and for the large-scale drug screening to target Ca2+ handling proteins.
KW - Calcium handling
KW - Calcium transient
KW - Cardiac disease modeling
KW - Cardiomyocyte maturation
KW - IPSCs
KW - Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs)
KW - Issue 155
KW - Medicine
UR - http://www.scopus.com/inward/record.url?scp=85078821473&partnerID=8YFLogxK
U2 - 10.3791/60135
DO - 10.3791/60135
M3 - Review article
C2 - 32009647
AN - SCOPUS:85078821473
SN - 1940-087X
VL - 2020
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 155
M1 - e60135
ER -