Yuan Qianliang, Maas Renee G C, Brouwer Ellen C J, Pei Jiayi, Blok Christian Snijders, Popovic Marko A, Paauw Nanne J, Bovenschen Niels, Hjortnaes Jesper, Harakalova Magdalena, Doevendans Pieter A, Sluijter Joost P G, van der Velden Jolanda, Buikema Jan W
Amsterdam Cardiovascular Sciences, Department of Physiology, Amsterdam University Medical Centers, De Boelelaan 1108, 1081 HZ Amsterdam, The Netherlands.
Utrecht Regenerative Medicine Center, Circulatory Health Laboratory, University Utrecht, 3584 CS Utrecht, The Netherlands.
J Cardiovasc Dev Dis. 2022 Jan 27;9(2):43. doi: 10.3390/jcdd9020043.
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 hiPSC-CMs 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.
成年心脏的收缩性与单个心肌细胞（CMs）中肌节的结构程度相关，并且似乎与再生能力呈负相关。在本研究中，我们利用多种成像技术追踪有丝分裂期间肌节解体的顺序，该过程导致源自人多能干细胞的增殖性心肌细胞（hiPSC-CMs）发生细胞或核分裂。我们观察到单核和双核hiPSC-CMs均可产生单核子细胞或双核后代。在这个用CHIR99021小分子处理的高度增殖性hiPSC-CMs来源中，我们发现与代谢成熟的非增殖性hiPSC-CMs和成人心脏组织相比，Wnt和Hippo信号更明显。此外，我们发现CHIR99021提高了高增殖性hiPSC-CMs中非病毒载体整合的效率，其中荧光转基因表达在染色体分离（M期）后出现。本研究为hiPSC-CMs和工程心脏组织中的基因操作研究提供了一种工具。此外，我们的数据表明，单核和双核CMs的细胞和核分裂背后存在复杂的生物学现象，有丝分裂期间肌节解体是一种共同现象。
