Massive expansion and cryopreservation of functional human induced pluripotent stem cell-derived cardiomyocytes

STAR Protoc. 2021 Feb 9;2(1):100334. doi: 10.1016/j.xpro.2021.100334. eCollection 2021 Mar 19.


Since the discovery of human induced pluripotent stem cells (hiPSCs), numerous strategies have been established to efficiently derive cardiomyocytes from hiPSCs (hiPSC-CMs). Here, we describe a cost-effective strategy for the subsequent massive expansion (>250-fold) of high-purity hiPSC-CMs relying on two aspects: removal of cell-cell contacts and small-molecule inhibition with CHIR99021. The protocol maintains CM functionality, allows cryopreservation, and the cells can be used in downstream assays such as disease modeling, drug and toxicity screening, and cell therapy. For complete details on the use and execution of this protocol, please refer to Buikema (2020).

Keywords: Cell culture; Cell differentiation; Stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Communication / drug effects*
  • Cryopreservation*
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism*
  • Pyridines / pharmacology*
  • Pyrimidines / pharmacology*


  • Chir 99021
  • Pyridines
  • Pyrimidines