Specific induction and long-term maintenance of high purity ventricular cardiomyocytes from human induced pluripotent stem cells

PLoS One. 2020 Nov 2;15(11):e0241287. doi: 10.1371/journal.pone.0241287. eCollection 2020.

Abstract

Currently, cardiomyocyte (CM) differentiation methods require a purification step after CM induction to ensure the high purity of the cell population. Here we show an improved human CM differentiation protocol with which high-purity ventricular-type CMs can be obtained and maintained without any CM purification process. We induced and collected a mesodermal cell population (platelet-derived growth factor receptor-α (PDGFRα)-positive cells) that can respond to CM differentiation cues, and then stimulated CM differentiation by means of Wnt inhibition. This method reproducibly generated CMs with purities above 95% in several human pluripotent stem cell lines. Furthermore, these CM populations were maintained in culture at such high purity without any further CM purification step for over 200 days. The majority of these CMs (>95%) exhibited a ventricular-like phenotype with a tendency to structural and electrophysiological maturation, including T-tubule-like structure formation and the ability to respond to QT prolongation drugs. This is a simple and valuable method to stably generate CM populations suitable for cardiac toxicology testing, disease modeling and regenerative medicine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation / genetics*
  • Cell Lineage / genetics
  • Electrophysiological Phenomena
  • Heart Ventricles / cytology
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Mesoderm / cytology
  • Mesoderm / growth & development*
  • Myocytes, Cardiac / cytology*
  • Receptor, Platelet-Derived Growth Factor alpha / genetics
  • Wnt Proteins / antagonists & inhibitors

Substances

  • Wnt Proteins
  • Receptor, Platelet-Derived Growth Factor alpha

Grants and funding

This research was supported by Japan Agency for Medical Research and Development (AMED) in the form of grants awarded to JKY (JP15mk0104053h0101; JP16mk0104027h0402; JP17mk0104027h0403) and Japan Society for the Promotion of Science (JSPS) in the form of a grant awarded to JKY (JP18H04048). The funder provided financial support, but did not have any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.