Engineered Heart Tissues for Contractile, Structural, and Transcriptional Assessment of Human Pluripotent Stem Cell-Derived Cardiomyocytes in a Three-Dimensional, Auxotonic Environment

Samantha Bremner; Alex J Goldstein; Ty Higashi; Nathan J Sniadecki

doi:10.1007/978-1-0716-2261-2_6

Engineered Heart Tissues for Contractile, Structural, and Transcriptional Assessment of Human Pluripotent Stem Cell-Derived Cardiomyocytes in a Three-Dimensional, Auxotonic Environment

Methods Mol Biol. 2022:2485:87-97. doi: 10.1007/978-1-0716-2261-2_6.

Authors

Samantha Bremner^#^{1

2}, Alex J Goldstein^#^{1

3

4}, Ty Higashi^#^{1

5}, Nathan J Sniadecki^{6

7

8

9}

Affiliations

¹ Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
² Department of Bioengineering, University of Washington, Seattle, WA, USA.
³ Department of Materials Science and Engineering, University of Washington, Seattle, WA, USA.
⁴ Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA, USA.
⁵ Department of Mechanical Engineering, University of Washington, Seattle, WA, USA.
⁶ Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA. nsniadec@uw.edu.
⁷ Department of Bioengineering, University of Washington, Seattle, WA, USA. nsniadec@uw.edu.
⁸ Department of Mechanical Engineering, University of Washington, Seattle, WA, USA. nsniadec@uw.edu.
⁹ Department of Laboratory Medicine & Pathology, University of Washington, Seattle, WA, USA. nsniadec@uw.edu.

^# Contributed equally.

PMID: 35618900
DOI: 10.1007/978-1-0716-2261-2_6

Abstract

Three-dimensional, human engineered heart tissue promotes maturation of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and provides a useful platform for in vitro cardiac development and disease modeling. This protocol describes the generation of fibrin-based engineered heart tissues (EHTs) containing hiPSC-CMs and human stromal cells. The platform makes use of racks of silicone posts that fit a standard 24-well dish. Stromal cells and hiPSC-CMs are cast in a fibrin hydrogel suspended between two silicone posts, forming an engineered tissue that generates synchronous contractions. The platform described herein is amenable to various measures of cardiac function including measurement of contractile force and calcium handling, as well as molecular biology assays and immunostaining.

Keywords: Cardiac tissue engineering; Cardiomyocytes; Engineered heart tissue; Fibrin; Induced pluripotent stem cells.

MeSH terms

Fibrin
Humans
Induced Pluripotent Stem Cells* / cytology
Myocytes, Cardiac* / cytology
Silicones
Tissue Engineering*

Substances

Silicones
Fibrin