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.
Novin Balafkan  1 ,  Sepideh Mostafavi  1 ,  Manja Schubert  2 ,  Richard Siller  3  4 ,  Kristina Xiao Liang  2 ,  Gareth Sullivan  5  3  4  6 ,  Laurence A Bindoff  7  8  9
Affiliations
  • 1. Department of Clinical Medicine (K1), University of Bergen, Bergen, Norway.
  • 2. Department of Neurology, Haukeland University Hospital, 5021, Bergen, Norway.
  • 3. Norwegian Center for Stem Cell Research, Oslo University Hospital, Domus Medica, Oslo, Norway.
  • 4. Institute of Immunology, Oslo University Hospital-Rikshospitalet, P.O. Box 4950, 0424, Nydalen, Oslo, Norway.
  • 5. Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
  • 6. Hybrid Technology Hub - Centre of Excellence, Institute of Basic Medical Sciences, University of Oslo, Blindern, Oslo, Norway.
  • 7. Department of Clinical Medicine (K1), University of Bergen, Bergen, Norway. [email protected].
  • 8. Department of Neurology, Haukeland University Hospital, 5021, Bergen, Norway. [email protected].
  • 9. Neuro-SysMed, Department of Neurology, Haukeland University Hospital, Bergen, Norway. [email protected].
PMID: 33116175 DOI: 10.1038/s41598-020-73656-2
Abstract

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.