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Updated: Oct 14, 2025

Generation of Human Cardiomyocytes: A Differentiation Protocol from Feeder-free Human Induced Pluripotent Stem Cells
Published on: June 28, 2013
A simple protocol to produce mature human-induced pluripotent stem cell-derived cardiomyocytes
Walter E Knight1,2,3, Yingqiong Cao1, Phoebe Dillon1,2,4
1Division of Cardiology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
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).

