Protocol detail

Cytotoxicity Assessment of Drugs on hiPSC-CMs

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iCell-Cardiomyocytes² (Fujifilm Cellular Dynamics Inc., Madison, WI, United States) were seeded into 96 well-plates (Greiner Bio One, ref. 655946) according to the manufacturer instructions, at a density of 50 k cells/well. Seven days after seeding, the hiPSC-CMs were used to assess cytotoxic effects of doxorubicin, captopril and sunitinib. To assess effects on viability and cytotoxicity, two assay combination kits were used: 1) RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay [Promega, JA1011 (Kupcho et al., 2019 (link))] and 2) ApoLive-Glo Multiplex Assay [Promega, G6410 (Flörkemeier et al., 2022 (link))]. The assays were performed according to manufacturer instructions. The experiments were repeated at 24, 48, and 72 h in order to assess any time-dependent change in live cells or activation of cell-death pathways.

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Altrocchi C., Van Ammel K., Steemans M., Kreir M., Tekle F., Teisman A., Gallacher D.J, & Lu H.R. (2023). Evaluation of chronic drug-induced electrophysiological and cytotoxic effects using human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Frontiers in Pharmacology, 14, 1229960.

Publication 2023

iCell-Cardiomyocytes2 RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay ApoLive-Glo Multiplex Assay

Annexin v Apoptosis Assays Captopril Cell death Cells Cytotoxicity Doxorubicin Hipsc Necrosis Promega Sunitinib

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Variable analysis

independent variables

Concentration of doxorubicin
Concentration of captopril
Concentration of sunitinib

dependent variables

Cell viability
Apoptosis
Necrosis

control variables

Cell type (iCell-Cardiomyocytes)
Cell seeding density (50 k cells/well)
Time points (24 h, 48 h, 72 h)

controls

Positive control: Not specified
Negative control: Not specified

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