Generation of Ventricular and Atrial-like hiPSC-CMs

hiPSC-CMs were generated from the control LUMC0099iCTRL04 hiPSC line, which was derived from human fibroblasts extracted through skin biopsies from of a Caucasian woman. The LUMC0099iCTRL04 line is registered in the Human Pluripotent Stem Cell Registry (Seltmann et al., 2016 (link)), which contains all details pertaining to its generation and characterization (hPSCreg, 2019 ). hiPSC clones showing stem cell morphology were characterized for pluripotency marker expression and differentiation potential to hiPSC-CMs in BPEL medium (Ng et al., 2008 (link)) containing activin-A, BMP4, and CHIR99021 (Devalla et al., 2016 (link)). After 3 days, this medium was replaced by BPEL medium containing XAV939 (Tocris Biosciences) for ventricular differentiation (Ng et al., 2008 (link); Devalla et al., 2016 (link)). To differentiate hiPSC-CMs to atrial-like hiPSC-CMs, 1 μM all-trans retinoic acid (RA) was added (Devalla et al., 2015 (link)). Twenty days after differentiation, hiPSC-CMs were dissociated with TrypLE Select (Life Technologies), and plated at a low density (≈7.5 × 10⁴ cells) on Matrigel coated coverslips in BPEL medium (Devalla et al., 2016 (link)).

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Hilderink S., Devalla H.D., Bosch L., Wilders R, & Verkerk A.O. (2020). Ultrarapid Delayed Rectifier K+ Channelopathies in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Frontiers in Cell and Developmental Biology, 8, 536.

Publication 2020

XAV939 TrypLE Select

Activin a All trans retinoic acid Atrial Biopsies Bmp4 Caucasian Cells Chir99021 Clones Fibroblasts Hipsc Human Matrigel Pluripotent stem cell Skin Stem Stem cell Ventricular Woman Xav939

Corresponding Organization : University of Amsterdam

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Variable analysis

independent variables

Activin-A, BMP4, and CHIR99021 in BPEL medium
XAV939 in BPEL medium
1 μM all-trans retinoic acid (RA)

dependent variables

Differentiation of hiPSC-CMs to ventricular and atrial-like phenotypes

control variables

LUMC0099iCTRL04 hiPSC line derived from human fibroblasts of a Caucasian woman
Characterization of hiPSC clones for pluripotency marker expression and differentiation potential

positive controls

BPEL medium containing activin-A, BMP4, and CHIR99021 for cardiac differentiation
BPEL medium containing XAV939 for ventricular differentiation
1 μM all-trans retinoic acid (RA) for atrial-like differentiation

negative controls

No negative controls explicitly mentioned

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