Sarcomere Disassembly and Transfection Efficiency in Proliferating Human iPSC-Derived Cardiomyocytes

Qianliang Yuan, Renee G.C. Maas, Ellen C.J. Brouwer, Jiayi Pei, Christian Snijders Blok, Marko A. Popovic, Nanne J. Paauw, Niels Bovenschen, Jesper Hjortnaes, Magdalena Harakalova, Pieter A. Doevendans, Joost P.G. Sluijter, Jolanda van der Velden, Jan W. Buikema*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Contractility of the adult heart relates to the architectural degree of sarcomeres in individual cardiomyocytes (CMs) and appears to be inversely correlated with the ability to regenerate. In this study we utilized multiple imaging techniques to follow the sequence of sarcomere disassembly during mitosis resulting in cellular or nuclear division in a source of proliferating human pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). We observed that both mono-and binuclear hiPSCCMs give rise to mononuclear daughter cells or binuclear progeny. Within this source of highly proliferative hiPSC-CMs, treated with the CHIR99021 small molecule, we found that Wnt and Hippo signaling was more present when compared to metabolic matured non-proliferative hiPSC-CMs and adult human heart tissue. Furthermore, we found that CHIR99021 increased the efficiency of non-viral vector incorporation in high-proliferative hiPSC-CMs, in which fluorescent transgene expression became present after the chromosomal segregation (M phase). This study provides a tool for gene manipulation studies in hiPSC-CMs and engineered cardiac tissue. Moreover, our data illustrate that there is a complex biology behind the cellular and nuclear division of mono-and binuclear CMs, with a shared-phenomenon of sarcomere disassembly during mitosis.

Original languageEnglish
Article number43
Pages (from-to)1-13
JournalJournal of Cardiovascular Development and Disease
Volume9
Issue number2
DOIs
Publication statusPublished - Feb 2022

Keywords

  • Binucleation
  • Cardiomyocyte proliferation
  • Cardiomyocytes
  • Human iPSC
  • IPSC-derived cardiomyocytes
  • M-phase
  • Mitosis
  • Non-viral vector incorporation
  • Proliferation
  • Sarcomere development
  • Sarcomere disassembly
  • Self-duplication
  • Transfection efficiency

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