Massive expansion and cryopreservation of functional human induced pluripotent stem cell-derived cardiomyocytes
Maas, Renee G.C.; Lee, Soah; Harakalova, Magdalena; Snijders Blok, Christian J.B.; Goodyer, William R.; Hjortnaes, Jesper; Doevendans, Pieter A.F.M.; Van Laake, Linda W.; van der Velden, Jolanda; Asselbergs, Folkert W.; Wu, Joseph C.; Sluijter, Joost P.G.; Wu, Sean M.; Buikema, Jan W.
(2021) STAR protocols, volume 2, issue 1
(Article)
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.
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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).
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Keywords: Cell culture, Cell differentiation, Stem cells, General Biochemistry,Genetics and Molecular Biology, General Neuroscience, General Immunology and Microbiology
ISSN: 2666-1667
Publisher: Cell Press
Note: Funding Information: This work was supported by a UMC Utrecht Clinical Fellowship, Netherlands Heart Institute Fellowship, and CVON-Dosis young talent grant (to J.W.B.); Stanford Child Health Research Institute Postdoctoral Fellowship and NIH NRSA Postdoctoral Fellowship 5F32HL142205 (to S.L.); R01 HL145676 , R01 HL146690 , and P01 HL141084 (to J.C.W); NIH ( OD004411 , HL099776 , LM012179 ) and the Joan and Sanford I. Weill Scholar Fund (to S.M.W.); Netherlands Heart Foundation (CVON-Dosis 2014–40), and Netherlands Organization for Sciences (NWO)-ZonMW (VICI 91818602) (to J.V.); Dutch Research Council (NWO) VENI grant no. 016.176.136 (M.H.); Foundation Leducq (Cure-PLaN) (to R.G.C.M., M.H., and F.A.). R.G.C.M. is supported by a grant of the PLN Foundation . F.W.A. is supported by UCL Hospitals NIHR Biomedical Research Centre ; Horizon2020 ERC-2016-COG EVICARE ( 725229 ), Horizon 2020 BRAV3 (SC1-BHC-07-2019) and ZonMw-TAS program (no. 116002016 ) (to J.S.). We thank Prof. Joseph Wu (University of Stanford, USA) for the provision of the SCVI-111, SCVI-114, and SCVI-273 hiPSC lines. The graphical abstract was created with BioRender.com. Funding Information: This work was supported by a UMC Utrecht Clinical Fellowship, Netherlands Heart Institute Fellowship, and CVON-Dosis young talent grant (to J.W.B.); Stanford Child Health Research Institute Postdoctoral Fellowship and NIH NRSA Postdoctoral Fellowship 5F32HL142205 (to S.L.); R01 HL145676, R01 HL146690, and P01 HL141084 (to J.C.W); NIH (OD004411, HL099776, LM012179) and the Joan and Sanford I. Weill Scholar Fund (to S.M.W.); Netherlands Heart Foundation (CVON-Dosis 2014–40), and Netherlands Organization for Sciences (NWO)-ZonMW (VICI 91818602) (to J.V.); Dutch Research Council (NWO) VENI grant no. 016.176.136 (M.H.); Foundation Leducq (Cure-PLaN) (to R.G.C.M. M.H. and F.A.). R.G.C.M. is supported by a grant of the PLN Foundation. F.W.A. is supported by UCL Hospitals NIHR Biomedical Research Centre; Horizon2020 ERC-2016-COG EVICARE (725229), Horizon 2020 BRAV3 (SC1-BHC-07-2019) and ZonMw-TAS program (no. 116002016) (to J.S.). We thank Prof. Joseph Wu (University of Stanford, USA) for the provision of the SCVI-111, SCVI-114, and SCVI-273 hiPSC lines. The graphical abstract was created with BioRender.com. R.G.C.M. designed and performed the experiments. R.G.C.M. S.L. and J.W.B. analyzed the data and wrote the manuscript. C.S.B. contributed to the development, analysis, and validation of the flow cytometry. J.P.G.S. S.M.W. and J.W.B. supervised the project. M.H. W.R.G. J.H. P.A.F.M.D. L.W.L. J.V. F.A. and J.C.W. edited and critically reviewed the manuscript. J.W.B. and S.M.W. have filed for a patent with the US Patent and Trademark Office regarding the effect of bioactive lipids plus Wnt signaling activation on hiPSC-CM proliferation/expansion. Publisher Copyright: © 2021 The Author(s)
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