Research Paper Volume 10, Issue 4 pp 532—548

A net-shaped multicellular formation facilitates the maturation of hPSC-derived cardiomyocytes through mechanical and electrophysiological stimuli

Taoyan Liu 1, 2, , Chengwu Huang 3, 4, , Hongxia Li 1, 2, , Fujian Wu 1, 2, , Jianwen Luo 3, 4, , Wenjing Lu 1, 2, , Feng Lan 1, 2, ,

  • 1 Beijing Laboratory for Cardiovascular Precision Medicine, The Key Laboratory of Remodeling-Related Cardiovascular Disease, Ministry of Education, Beijing Collaborative Innovation Center for Cardiovascular Disorders, Anzhen Hospital, Capital Medical University, Beijing 100029, China
  • 2 Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing 100029, China
  • 3 Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084, China
  • 4 Center for Biomedical Imaging Research, Tsinghua University, Beijing 100084, China

received: February 4, 2020 ; accepted: April 9, 2018 ; published: April 14, 2018 ;

https://doi.org/10.18632/aging.101411
How to Cite

Copyright: Liu et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

The use of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is limited in drug discovery and cardiac disease mechanism studies due to cell immaturity. Although many approaches have been reported to improve the maturation of hiPSC-CMs, the elucidation of the process of maturation is crucial. We applied a small-molecule-based differentiation method to generate cardiomyocytes (CMs) with multiple aggregation forms. The motion analysis revealed significant physical differences in the differently shaped CMs, and the net-shaped CMs had larger motion amplitudes and faster velocities than the sheet-shaped CMs. The net-shaped CMs displayed accelerated maturation at the transcriptional level and were more similar to CMs with a prolonged culture time (30 days) than to sheet-d15. Ion channel genes and gap junction proteins were up-regulated in net-shaped CMs, indicating that robust contraction was coupled with enhanced ion channel and connexin expression. The net-shaped CMs also displayed improved myofibril ultrastructure under transmission electron microscopy. In conclusion, different multicellular hPSC-CM structures, such as the net-shaped pattern, are formed using the conditioned induction method, providing a useful tool to improve cardiac maturation.

Abbreviations

CM: cardiomyocyte; hiPSC: human induced-pluripotent stem cell; HCM: hypertrophic cardiomyopathy; DCM: dilated cardiomyopathy; LQT: long QT syndrome; cTnI: cardiac troponin I; cTnT: cardiac troponin T; CX43: connexin-43; DAPI: 4,6-diamidino-2-phenylindole; qRT-PCR: quantitative real-time polymerase chain reaction; MHMA: maximum horizontal motion amplitude; MLMA: maximum longitudinal motion amplitude; MV: maximum velocity; MHV: maximum horizontal velocity; MLV: maximum longitudinal velocity; LRRC39: leucine-rich repeat-containing protein 39; SEARCA2: sarco/endoplasmic reticulum Ca2+ ATPase RYR2: ryanodine receptor 2; ECM: extracellular matrix; EDTA: ethylenediaminetetraacetic acid; GFP: green fluorescent protein; BSA: bovine serum albumin; PBS: phosphate-buffered saline.