Large-scale production of cardiomyocytes from human pluripotent stem cells using a highly reproducible small molecule-based differentiation protocol

Hananeh Fonoudi, Hassan Ansari, Saeed Abbasalizadeh, Gillian M. Blue, Nasser Aghdami, David S. Winlaw, Richard P. Harvey, Alexis Bosman*, Hossein Baharvand

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Maximizing the benefit of human pluripotent stem cells (hPSCs) for research, disease modeling, pharmaceutical and clinical applications requires robust methods for the large-scale production of functional cell types, including cardiomyocytes. Here we demonstrate that the temporal manipulation of WNT, TGF-β, and SHH signaling pathways leads to highly efficient cardiomyocyte differentiation of single-cell passaged hPSC lines in both static suspension and stirred suspension bioreactor systems. Employing this strategy resulted in ~ 100% beating spheroids, consistently containing > 80% cardiac troponin T-positive cells after 15 days of culture, validated in multiple hPSC lines. We also report on a variation of this protocol for use with cell lines not currently adapted to single-cell passaging, the success of which has been verified in 42 hPSC lines. Cardiomyocytes generated using these protocols express lineage-specific markers and show expected electrophysiological functionalities. Our protocol presents a simple, efficient and robust platform for the large-scale production of human cardiomyocytes.

Original languageEnglish (US)
Article numbere54276
JournalJournal of Visualized Experiments
Volume2016
Issue number113
DOIs
StatePublished - Jul 25 2016

Keywords

  • Cardiac differentiation
  • Cardiomyocytes
  • Developmental Biology
  • Human pluripotent stem cells
  • Issue 113
  • Small molecules
  • Stem cell biology
  • Stirred suspension bioreactor

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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