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Review
. 2022 Aug 18;3(3):101560.
doi: 10.1016/j.xpro.2022.101560. eCollection 2022 Sep 16.

A review of protocols for human iPSC culture, cardiac differentiation, subtype-specification, maturation, and direct reprogramming

Davi M Lyra-Leite  1 , Óscar Gutiérrez-Gutiérrez  2 , Meimei Wang  3 , Yang Zhou  3 , Lukas Cyganek  2 , Paul W Burridge  4
Affiliations
Review

A review of protocols for human iPSC culture, cardiac differentiation, subtype-specification, maturation, and direct reprogramming

Davi M Lyra-Leite et al. STAR Protoc. .

Abstract

The methods for the culture and cardiomyocyte differentiation of human embryonic stem cells, and later human induced pluripotent stem cells (hiPSC), have moved from a complex and uncontrolled systems to simplified and relatively robust protocols, using the knowledge and cues gathered at each step. HiPSC-derived cardiomyocytes have proven to be a useful tool in human disease modelling, drug discovery, developmental biology, and regenerative medicine. In this protocol review, we will highlight the evolution of protocols associated with hPSC culture, cardiomyocyte differentiation, sub-type specification, and cardiomyocyte maturation. We also discuss protocols for somatic cell direct reprogramming to cardiomyocyte-like cells.

Keywords: Cell Differentiation; Cell culture; Stem Cells.

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Figures

Figure 1
Figure 1
hPSC culture, differentiation, and applications (A) hiPSC are cultured on Matrigel or Synthemax using chemically defined media until differentiation, with continuous monitoring of morphology and confluence levels. Before differentiation, hiPSC can also be replated or aggregated depending on the differentiation protocol. (B) Examples of differentiation protocols to obtain ventricular (also Table 1), atrial, and nodal (Table 2) hiPSC-CM. (C) Further culture approaches and applications of hiPSC-CM using protocols described in this paper.
Figure 2
Figure 2
Direct reprogramming of CFs into induced cardiomyocytes In vivo reprogramming is performed via injection of transcription factors into the injured area of the heart that will lead to de novo cardiomyocyte formation. In vitro reprogramming is performed via treatment of (cardiac) fibroblasts with different cocktails to generate iCMs. These cells can later be re-transplanted into the heart if needed to recover injured areas.
Figure 3
Figure 3
Different maturation approaches for hPSC-CMs Mechanical, chemical, and cellular-based cues can be used to induced maturation of hPSC-CMs. Each method has the potential to change to different degrees the morphology, metabolism, physiology, and gene expression of cardiomyocytes.
Figure 4
Figure 4
Tissue engineered approaches for further culture of hPSC-CMs Post-differentiation hPSC-CMs can be engineered into different types of constructs for disease modeling, pharmacological screenings, and developmental studies. Furthermore, each engineered construct has the potential to promote phenotypical changes in the hPSC-CMs, as listed here.
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