Single-cell RNA sequencing reveals maturation trajectory in human pluripotent stem cell-derived cardiomyocytes in engineered tissues

Shangli Cheng; David Brenière-Letuffe; Virpi Ahola; Andy O T Wong; Hoi Yee Keung; Bimal Gurung; Zongli Zheng; Kevin D Costa; Deborah K Lieu; Wendy Keung; Ronald A Li

doi:10.1016/j.isci.2023.106302

Single-cell RNA sequencing reveals maturation trajectory in human pluripotent stem cell-derived cardiomyocytes in engineered tissues

iScience. 2023 Mar 1;26(4):106302. doi: 10.1016/j.isci.2023.106302. eCollection 2023 Apr 21.

Authors

Shangli Cheng¹, David Brenière-Letuffe^{1

2

3}, Virpi Ahola¹, Andy O T Wong⁴, Hoi Yee Keung¹, Bimal Gurung^{4

5}, Zongli Zheng¹, Kevin D Costa^{4

6}, Deborah K Lieu^{4

7}, Wendy Keung^{4

8}, Ronald A Li⁴

Affiliations

¹ Ming Wai Lau Centre for Reparative Medicine, Karolinska Institutet, Hong Kong SAR, China.
² Department of Clinical Sciences, Intervention and Technology, CLINTEC, Karolinska Institutet, 141 52 Stockholm, Sweden.
³ Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, 141 86 Stockholm, Sweden.
⁴ Novoheart, Irvine, CA 92617, USA.
⁵ School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR, China.
⁶ Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁷ Institute for Regenerative Cures and Stem Cell Program, University of California, Davis, Sacramento, CA 95817, USA.
⁸ Dr. Li Dak Sum Research Centre, The University of Hong Kong, Hong Kong SAR, China.

Abstract

Cardiac in vitro models have become increasingly obtainable and affordable with the optimization of human pluripotent stem cell-derived cardiomyocyte (hPSC-CM) differentiation. However, these CMs are immature compared to their in vivo counterparts. Here we study the cellular phenotype of hPSC-CMs by comparing their single-cell gene expression and functional profiles in three engineered cardiac tissue configurations: human ventricular (hv) cardiac anisotropic sheet, cardiac tissue strip, and cardiac organoid chamber (hvCOC), with spontaneously aggregated 3D cardiac spheroids (CS) as control. The CM maturity was found to increase with increasing levels of complexity of the engineered tissues from CS to hvCOC. The contractile components are the first function to mature, followed by electrophysiology and oxidative metabolism. Notably, the 2D tissue constructs show a higher cellular organization whereas metabolic maturity preferentially increases in the 3D constructs. We conclude that the tissue engineering models resembling configurations of native tissues may be reliable for drug screening or disease modeling.

Keywords: Cell biology; Stem cells research; Tissue Engineering.