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

Renee G C Maas; Soah Lee; Magdalena Harakalova; Christian J B Snijders Blok; William R Goodyer; Jesper Hjortnaes; Pieter A F M Doevendans; Linda W Van Laake; Jolanda van der Velden; Folkert W Asselbergs; Joseph C Wu; Joost P G Sluijter; Sean M Wu; Jan W Buikema

doi:10.1016/j.xpro.2021.100334

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.

Authors

Renee G C Maas¹, Soah Lee², Magdalena Harakalova¹, Christian J B Snijders Blok¹, William R Goodyer², Jesper Hjortnaes³, Pieter A F M Doevendans¹, Linda W Van Laake¹, Jolanda van der Velden⁴, Folkert W Asselbergs^{5

6

7}, Joseph C Wu², Joost P G Sluijter¹, Sean M Wu², Jan W Buikema^{1

2}

Affiliations

¹ Utrecht Regenerative Medicine Center, Circulatory Health Laboratory, University Utrecht, Department of Cardiology, University Medical Center Utrecht, 3508 GA Utrecht, the Netherlands.
² Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA.
³ Department of Cardiothoracic Surgery, Heart & Lung Center, Leiden University Medical Center, Leiden, the Netherlands.
⁴ Amsterdam Cardiovascular Sciences, Department of Physiology, Amsterdam University Medical Centers, Amsterdam, the Netherlands.
⁵ Department of Cardiology, Division Heart & Lungs, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands.
⁶ Institute of Cardiovascular Science, Faculty of Population Health Sciences, University College London, London, UK.
⁷ Health Data Research UK and Institute of Health Informatics, University College London, London, UK.

Abstract

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*

Substances

Chir 99021
Pyridines
Pyrimidines

Abstract

Publication types

MeSH terms

Substances

Grants and funding