Engineered Heart Tissues for Standard 96-Well Tissue Culture Plates

Alex J Goldstein; Ruby M Padgett; Samantha B Bremner; Ty Higashi; Ava M Obenaus; Sam Bolduan; David L Mack; Nathan J Sniadecki

doi:10.1007/978-1-0716-3854-5_5

Engineered Heart Tissues for Standard 96-Well Tissue Culture Plates

Methods Mol Biol. 2024:2805:89-100. doi: 10.1007/978-1-0716-3854-5_5.

Authors

Alex J Goldstein^{1

2

3}, Ruby M Padgett^{2

3

4}, Samantha B Bremner^{2

3

5}, Ty Higashi^{2

3

4}, Ava M Obenaus^{2

3

4}, Sam Bolduan⁴, David L Mack^{2

3

5

6}, Nathan J Sniadecki^{7

8

9

10

11}

Affiliations

¹ Department of Lab Medicine and Pathology, University of Washington, Seattle, WA, USA.
² Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA.
³ Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
⁴ Department of Mechanical Engineering, University of Washington, Seattle, WA, USA.
⁵ Department of Bioengineering, University of Washington, Seattle, WA, USA.
⁶ Department of Rehabilitation Medicine, University of Washington, Seattle, WA, USA.
⁷ Department of Lab Medicine and Pathology, University of Washington, Seattle, WA, USA. [email protected].
⁸ Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA. [email protected].
⁹ Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA. [email protected].
¹⁰ Department of Mechanical Engineering, University of Washington, Seattle, WA, USA. [email protected].
¹¹ Department of Bioengineering, University of Washington, Seattle, WA, USA. [email protected].

PMID: 39008175
DOI: 10.1007/978-1-0716-3854-5_5

Abstract

Engineered heart tissues (EHTs) have been shown to be a valuable platform for disease investigation and therapeutic testing by increasing human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) maturity and better recreating the native cardiac environment. The protocol detailed in this chapter describes the generation of miniaturized EHTs (mEHTs) incorporating hiPSC-CMs and human stromal cells in a fibrin hydrogel. This platform utilizes an array of silicone posts designed to fit in a standard 96-well tissue culture plate. Stromal cells and hiPSC-CMs are cast in a fibrin matrix suspended between two silicone posts, forming an mEHT that produces synchronous muscle contractions. The platform presented here has the potential to be used for high throughput characterization and screening of disease phenotypes and novel therapeutics through measurements of the myocardial function, including contractile force and calcium handling, and its compatibility with immunostaining.

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

MeSH terms

Cell Culture Techniques / methods
Cell Differentiation
Cells, Cultured
Fibrin / metabolism
Humans
Hydrogels / chemistry
Induced Pluripotent Stem Cells* / cytology
Induced Pluripotent Stem Cells* / metabolism
Myocytes, Cardiac* / cytology
Myocytes, Cardiac* / metabolism
Stromal Cells / cytology
Tissue Culture Techniques / instrumentation
Tissue Culture Techniques / methods
Tissue Engineering* / methods

Substances

Hydrogels
Fibrin