Abstract
The immature phenotype of human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) is a major limitation to the use of these valuable cells for pre-clinical toxicity testing and for disease modeling. Here we tested the hypothesis that human perinatal stem cell derived extracellular matrix (ECM) promotes hiPSC-CM maturation to a greater extent than mouse cell derived ECM. We refer to the human ECM as Matrix Plus (Matrix Plus) and compare effects to commercially available mouse ECM (Matrigel). hiPSC-CMs cultured on Matrix Plus mature functionally and structurally seven days after thaw from cryopreservation. Mature hiPSC-CMs showed rod-shaped morphology, highly organized sarcomeres, elevated cTnI expression and mitochondrial distribution and function like adult cardiomyocytes. Matrix Plus also promoted mature hiPSC-CM electrophysiological function and monolayers’ response to hERG ion channel specific blocker was Torsades de Pointes (TdP) reentrant arrhythmia activations in 100% of tested monolayers. Importantly, Matrix Plus enabled high throughput cardiotoxicity screening using mature human cardiomyocytes with validation utilizing reference compounds recommended for the evolving Comprehensive In Vitro Proarrhythmia Assay (CiPA) coordinated by the Health and Environmental Sciences Institute (HESI). Matrix Plus offers a solution to the commonly encountered problem of hiPSC-CM immaturity that has hindered implementation of these human based cell assays for pre-clinical drug discovery.
Highlights
The immature phenotype of human induced pluripotent stem cell derived cardiomyocytes is a major limitation to the use of these valuable cells for pre-clinical toxicity testing and for disease modeling
Matrix Plus was generated from human perinatal stem cells using methods similar to those established with other matrix-producing cells[27]
The cardiomyocyte maturation phenomenon present on Matrix Plus is not observed on CELLvo Matrix, suggesting that any differences are potentially related to the mechanism of action driving the maturation process
Summary
The immature phenotype of human induced pluripotent stem cell derived cardiomyocytes (hiPSCCMs) is a major limitation to the use of these valuable cells for pre-clinical toxicity testing and for disease modeling. Current approaches to hiPSC-CM maturation include: long term culture of at least 30–100 days[10], exercising cells mechanically and e lectrically[11,12], metabolic and hormonal manipulations[13], co-culture with other cell types14, 3D organoid development[15,16] and modulation of extracellular matrix (ECM) properties[17] Of these approaches ECM manipulation in 2D hiPSC-CM monolayers offers a highly feasible option with quantifiable variables that are compatible with high throughput screening assays. We present Matrix Plus-a novel human in vitro perinatal stem cell-derived ECM that supports rapid (7 days) structural and functional maturation of hiPSC-CM phenotypes in 2D monolayer culture. The utility of Matrix Plus coated 96 well plates for hiPSC-CM proarrhythmia assays is validated using clinically relevant doses of medications with known risk to cause arrhythmias as outlined in the recent hiPSC-CM validation assay
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