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. 2018 Oct:32:65-72.
doi: 10.1016/j.scr.2018.08.015. Epub 2018 Aug 24.

A complete workflow for the differentiation and the dissociation of hiPSC-derived cardiospheres

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Free article

A complete workflow for the differentiation and the dissociation of hiPSC-derived cardiospheres

Benjamin Fischer et al. Stem Cell Res. 2018 Oct.
Free article

Abstract

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) are an invaluable tool for both basic and translational cardiovascular research. The potential that these cells hold for therapy, disease modeling and drug discovery is hampered by several bottlenecks that currently limit both the yield and the efficiency of cardiac induction. Here, we present a complete workflow for the production of ready-to-use hiPSC-CMs in a dynamic suspension bioreactor. This includes the efficient and highly reproducible differentiation of hiPSCs into cardiospheres, which display enhanced physiological maturation compared to static 3D induction in hanging drops, and a novel papain-based dissociation method that offers higher yield and viability than the broadly used dissociation reagents TrypLE and Accutase. Molecular and functional analyses of the cardiomyocytes reseeded after dissociation confirmed both the identity and the functionality of the cells, which can be used in downstream applications, either as monolayers or spheroids.

Keywords: 3D bioreactor; Cardiac induction; Cardiomyocytes; Papain dissociation; hiPSCs.

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