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  • A simple protocol to produce mature human-induced pluripotent stem cell-derived cardiomyocytes.

A simple protocol to produce mature human-induced pluripotent stem cell-derived cardiomyocytes.

STAR protocols (2021-11-11)
Walter E Knight, Yingqiong Cao, Phoebe Dillon, Kunhua Song
ABSTRACT

When cultured under typical conditions, human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are structurally and functionally immature. We have previously demonstrated that culture of hiPSC-CMs in maturation medium containing fatty acids, in combination with culture on micropatterned surfaces, produces cells that demonstrate a more mature phenotype compared to standard approaches. Here, we show in detail the steps needed to produce mature hiPSC-CMs. Compared with many approaches, our protocol is relatively simple and can be easily adapted to new laboratories. For complete details on the use and execution of this protocol, please refer to Knight et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MOPS, ≥99.5% (titration)
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, ≥97.0%
Millipore
EDTA, ULTROL® Grade, 500 mM Solution, pH 8.0
Sigma-Aldrich
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
Monoclonal Anti-α-Actinin (Sarcomeric) antibody produced in mouse, clone EA-53, ascites fluid
Sigma-Aldrich
Urea, ACS reagent, 99.0-100.5%
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Gelatin from porcine skin, gel strength 300, Type A
Sigma-Aldrich
Potassium hydroxide, BioXtra, ≥85% KOH basis
Sigma-Aldrich
Sucrose, for molecular biology, ≥99.5% (GC)