Retinoic acid modulation guides human-induced pluripotent stem cell differentiation towards left or right ventricle-like cardiomyocytes

Hengliang Zhang ^1,2,3, Payel Sen ^1,2,4, Jules Hamers ^1,2,4

¹Walter Brendel Center for Experimental Medicine (WBex), University Clinic Munich, LMU Munich, 81377, Munich, Germany.
²Center for Cardiovascular Research (DZHK), Munich Heart Alliance (MHA), Partner Site Munich, 81377, Munich, Germany.
³The First Affiliated Hospital, College of Clinical Medicine of Henan, University of Science and Technology, Luoyang, China.
⁴Interfaculty Center for Endocrine and Cardiovascular Disease Network Modelling and Clinical Transfer (ICONLMU), LMU Munich, Munich, Germany.
⁵First Department of Medicine, Klinikum Rechts der Isar, School of Medicine and Health, Technical University of Munich, Cardiology, Munich, Germany.
⁶Regenerative Medicine in Cardiovascular Diseases, First Department of Medicine, Klinikum Rechts der Isar, School of Medicine and Health, Technical University of Munich, Munich, Germany.
⁷Walter Brendel Center for Experimental Medicine (WBex), University Clinic Munich, LMU Munich, 81377, Munich, Germany. daphne.merkus@med.uni-muenchen.de.
⁸Center for Cardiovascular Research (DZHK), Munich Heart Alliance (MHA), Partner Site Munich, 81377, Munich, Germany. daphne.merkus@med.uni-muenchen.de.
⁹Interfaculty Center for Endocrine and Cardiovascular Disease Network Modelling and Clinical Transfer (ICONLMU), LMU Munich, Munich, Germany. daphne.merkus@med.uni-muenchen.de.
¹⁰Division of Experimental Cardiology, Dept of Cardiology, Erasmus University Medical Center, 3000CA, Rotterdam, The Netherlands. daphne.merkus@med.uni-muenchen.de.

⁰Walter Brendel Center for Experimental Medicine (WBex), University Clinic Munich, LMU Munich, 81377, Munich, Germany.

Stem Cell Research & Therapy +

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June 20, 2024

View abstract on PubMed

Summary

Retinoic acid (RA) treatment of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) successfully generated cells and engineered heart tissues (EHTs) with a left or right ventricular phenotype. This method enhances cardiac cell differentiation and function for regenerative medicine applications.

Area Of Science

Cardiology
Stem Cell Biology
Developmental Biology

Background

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) traditionally yield mixed cell populations.
Retinoic acid (RA) is crucial for embryonic heart development.
Standard hiPSC-CM differentiation lacks specific chamber phenotypes.

Purpose Of The Study

To investigate the effect of retinoic acid (RA) on hiPSC-CM differentiation.
To generate hiPSC-CMs and engineered heart tissues (EHTs) with specific ventricular phenotypes.
To optimize hiPSC-CM differentiation for cardiac regenerative medicine.

Main Methods

hiPSC-CMs were differentiated with varying RA concentrations (0, 0.05μM, 0.1 μM) between days 3-6.
Engineered heart tissues (EHTs) were created by assembling hiPSC-CMs in a collagen hydrogel.
Cardiac marker gene expression, EHT contractility, and sensitivity to hypoxia/isoprenaline were analyzed.

Main Results

High RA concentration (HRA) group showed increased expression of contractile proteins (MYH6, MYH7, cTnT) and ventricular markers (TBX5, NKX2.5, CORIN).
HRA-derived EHTs exhibited higher contraction force, lower beating frequency, and increased sensitivity to hypoxia and isoprenaline, mimicking left ventricular function.
RNA sequencing indicated RA promotes extracellular matrix strength, enhancing EHT contractility.

Conclusions

Specific RA concentrations and timing during hiPSC differentiation can induce CMs and EHTs with distinct left or right ventricular phenotypes.
This targeted approach improves the functional characteristics of hiPSC-derived cardiac tissues.
The findings offer a method to generate chamber-specific cardiomyocytes for cardiac repair strategies.

Keywords:

Cardiomyocyte Engineered heart tissue Left ventricle Retinoic acid hiPSC