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Stanford University School of Medicine

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Cell Stem Cell, Volume 27, Issue 1, 2 July 2020, Pages 50-63.e5 | https://doi.org/10.1016/j.stem.2020.06.001 Wnt Activation and Reduced Cell-Cell Contact Synergistically Induce Massive Expansion of Functional Human iPSC-Derived Cardiomyocytes

Jan W. Buikema^1,2,16, Soah Lee^1,16, William R. Goodyer^1,3,17, Renee G. Maas^2,17, Orlando Chirikian^1,¹⁷, Guang Li¹, Yi Miao⁵, Sharon L. Paige^1,3, Daniel Lee¹, Haodi Wu¹, David T. Paik¹, Siyeon Rhee⁴, Lei Tian¹, Francisco X. Galdos¹, Nazan Puluca¹, Benjamin Beyersdorf¹, James Hu¹, Aimee Beck¹, Sneha Venkamatran¹, Srilatha Swami⁶, Paul Wijnker⁷, Maike Schuldt⁷, Larissa M. Dorsch⁷, Alain van Mil², Kristy Red-Horse^1,4, Joy Y. Wu⁶, Caroline Geisen⁸, Michael Hesse⁸, Vahid Serpooshan^1,9, Stefan Jovinge^10,11, Bernd K. Fleischmann⁸, Pieter A. Doevendans^2,12, Jolanda van der Velden⁷, K. Christopher Garcia⁵, Joseph C. Wu^1,13,14,15, Joost P.G. Sluijter², Sean M. Wu^1,14,15,18,*

¹Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA 94305, USA
²Utrecht Regenerative Medicine Center, Circulatory Health Laboratory, University Utrecht, Department of Cardiology, University Medical Center Utrecht, 3508 GA Utrecht, the Netherlands
³Division of Pediatric Cardiology, Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA
⁴Department of Biology, Stanford University, Stanford, CA 94305, USA
⁵Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA
⁶Division of Endocrinology and Metabolism, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
⁷Amsterdam Cardiovascular Sciences, Department of Physiology, Amsterdam University Medical Centers, Amsterdam, the Netherlands
⁸Institute of Physiology I, Life and Brain Center, Medical Faculty, University of Bonn, 53105 Bonn, Germany
⁹Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA 30322, USA
¹⁰DeVos Cardiovascular Research Program of Spectrum Health and Van Andel Research Institute, 100 Michigan Street NE, Grand Rapids, MI 49503, USA
¹¹Michigan State University, College of Human Medicine, 15 Michigan Street NE, Grand Rapids, MI, USA
¹²Netherlands Heart Institute, Holland Heart House, 3511 EP Utrecht, the Netherlands
¹³Department of Radiology, Stanford University School of Medicine, Stanford, CA 94305, USA
¹⁴Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
¹⁵Division of Cardiovascular Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
¹⁶These authors contributed equally
¹⁷These authors contributed equally
¹⁸Lead Contact

^*Corresponding author

Abstract

Modulating signaling pathways including Wnt and Hippo can induce cardiomyocyte proliferation in vivo. Applying these signaling modulators to human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in vitro can expand CMs modestly (<5-fold). Here, we demonstrate massive expansion of hiPSC-CMs in vitro (i.e., 100- to 250-fold) by glycogen synthase kinase-3β (GSK-3β) inhibition using CHIR99021 and concurrent removal of cell-cell contact. We show that GSK-3β inhibition suppresses CM maturation, while contact removal prevents CMs from cell cycle exit. Remarkably, contact removal enabled 10 to 25 times greater expansion beyond GSK-3β inhibition alone. Mechanistically, persistent CM proliferation required both LEF/TCF activity and AKT phosphorylation but was independent from yes-associated protein (YAP) signaling. Engineered heart tissues from expanded hiPSC-CMs showed comparable contractility to those from unexpanded hiPSC-CMs, demonstrating uncompromised cellular functionality after expansion. In summary, we uncovered a molecular interplay that enables massive hiPSC-CM expansion for large-scale drug screening and tissue engineering applications.

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형식Research article
게재일2020년 07월 (BRIC 등록일 2020-07-07)
연구진국외 연구진
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