104 Chapter 4 Problem 8: The hiPSC-CMs are low in their proliferation rate during passaging. Potential Solution: Optimize the CHIR concentration and add the CHIR every other day. Do not re-freeze CHIR aliquots. Use a lower splitting ratio in the first 3 passages (1:20-1:5) and increase the splitting ratio upon passage 3 and passage 4 to 1:2. Do not split after passage 4. Only use differentiated cell populations with ≥70% α-actinin-positive cells. ACKNOWLEGEMENTS 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 (SC1BHC-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.
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