Methods in Molecular Biology (2022) 2436: 67–81

DOI 10.1007/7651_2021_423

© Springer Science+Business Media, LLC 2021

Published online: 15 September 2021

Integrating Human-Induced Pluripotent Stem Cell

Expansion Capability and Cardiomyocyte Differentiation

Potential in a Microcarrier Suspension Culture

Valerie Ho, Gerine Tong, Alan Lam, Shaul Reuveny, and Steve Oh

Abstract

Human-induced pluripotent stem cells are known for their high proliferation capacity as well as their ability

to differentiate to different lineages (Ban et al., Theranostics 7(7):2067–2077, 2017; Chen et al., Stem Cell

Res 15(2):365–375, 2015; Serra et al., Trends Biotechnol 30(6):350–359, 2012). For stem-cell-derived

cardiomyocytes to evolve into a scalable therapeutic source, a large quantity of highly pure cardiomyocytes

is needed. Thus, lies the challenge of defining an efficient cardiomyocyte differentiation process. This

chapter describes a method to evaluate multiple human-induced pluripotent stem cell lines for their cardiac

differentiation potentials before evaluating their integrated proliferation and differentiation abilities in

microcarrier cultures in a spinner culture format.

Key words Cardiomyocytes, Human-induced pluripotent stem cells, Microcarrier

1

Introduction

Cardiac-related diseases are the one of the leading causes of death

around the world [1]. Since mature cardiomyocytes (CM) are

unable to regenerate to restore original functionality, the only way

to restore a damaged heart is through heart transplantation

[1, 2]. However, demand for such treatment exceeds the supply

due to the shortage of donors [1, 2]. To satisfy the demand, stem-

cell-derived CMs can be considered as a potential solution to

replace damaged cardiac tissue. For this reason, an optimized,

scalable, efficient system of producing high-purity stem-cell-

derived CMs utilizing microcarriers (MC) is needed [3]. By taking

advantage of human-induced pluripotent stem cells’ high prolifera-

tion as well as their ability to differentiate into CMs, the vision for a

renewable source of mature CMs may be realized [4–6]. Different

cell lines are shown to have different cardiac differentiation poten-

tials as well as growth rates on microcarriers [7]. The first step is

cell-line selection for high cardiac differentiation potentials on the

monolayer cultures. While the second is selection for high prolifer-

ation capacity in an MC spinner culture followed by differentiation

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