A method for differentiating human induced pluripotent stem cells toward functional cardiomyocytes in 96-well microplates

Sci Rep. 2020 Oct 28;10(1):18498. doi: 10.1038/s41598-020-73656-2.


The capacity of pluripotent stem cells both for self-renewal and to differentiate into any cell type have made them a powerful tool for studying human disease. Protocols for efficient differentiation towards cardiomyocytes using defined, serum-free culture medium combined with small molecules have been developed, but thus far, limited to larger formats. We adapted protocols for differentiating human pluripotent stem cells to functional human cardiomyocytes in a 96-well microplate format. The resulting cardiomyocytes expressed cardiac specific markers at the transcriptional and protein levels and had the electrophysiological properties that confirmed the presence of functional cardiomyocytes. We suggest that this protocol provides an incremental improvement and one that reduces the impact of heterogeneity by increasing inter-experimental replicates. We believe that this technique will improve the applicability of these cells for use in developmental biology and mechanistic studies of disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Differentiation*
  • Cell Line
  • Cell Separation / methods*
  • Culture Media, Serum-Free
  • Electrophysiological Phenomena / drug effects
  • Flow Cytometry
  • Humans
  • Myocardium / cytology
  • Myocytes, Cardiac / cytology*
  • Pluripotent Stem Cells / cytology*
  • Reproducibility of Results


  • Culture Media, Serum-Free