3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes

Cláudia Correia, Alexey Koshkin, Patrícia Duarte, Dongjian Hu, Madalena Carido, Maria J. Sebastião, Patrícia Gomes-Alves, David A. Elliott, Ibrahim J. Domian, Ana Palma Teixeira, Paula M. Alves, Margarida Serra

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Three-dimensional (3D) cultures of human pluripotent stem cell derived cardiomyocytes (hPSC-CMs) hold great promise for drug discovery, providing a better approximation to the in vivo physiology over standard two-dimensional (2D) monolayer cultures. However, the transition of CM differentiation protocols from 2D to 3D cultures is not straightforward. In this work, we relied on the aggregation of hPSC-derived cardiac progenitors and their culture under agitated conditions to generate highly pure cardiomyocyte aggregates. Whole-transcriptome analysis and 13C-metabolic flux analysis allowed to demonstrate at both molecular and fluxome levels that such 3D culture environment enhances metabolic maturation of hiPSC-CMs. When compared to 2D, 3D cultures of hiPSC-CMs displayed down-regulation of genes involved in glycolysis and lipid biosynthesis and increased expression of genes involved in OXPHOS. Accordingly, 3D cultures of hiPSC-CMs had lower fluxes through glycolysis and fatty acid synthesis and increased TCA-cycle activity. Importantly, we demonstrated that the 3D culture environment reproducibly improved both CM purity and metabolic maturation across different hPSC lines, thereby providing a robust strategy to derive enriched hPSC-CMs with metabolic features closer to that of adult CMs.

Original languageEnglish
Pages (from-to)630-644
Number of pages15
JournalBiotechnology and Bioengineering
Volume115
Issue number3
DOIs
Publication statusPublished - 1 Mar 2018

Fingerprint

Induced Pluripotent Stem Cells
Pluripotent Stem Cells
Stem cells
Cell culture
Cardiac Myocytes
Glycolysis
Genes
Metabolic Flux Analysis
Fluxes
Activity Cycles
Biosynthesis
Physiology
Gene Expression Profiling
Drug Discovery
Fatty acids
Lipids
Monolayers
Fatty Acids
Down-Regulation
Agglomeration

Keywords

  • 3D aggregates
  • fluxome
  • human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs)
  • metabolic maturation
  • transcriptome

Cite this

Correia, Cláudia ; Koshkin, Alexey ; Duarte, Patrícia ; Hu, Dongjian ; Carido, Madalena ; Sebastião, Maria J. ; Gomes-Alves, Patrícia ; Elliott, David A. ; Domian, Ibrahim J. ; Teixeira, Ana Palma ; Alves, Paula M. ; Serra, Margarida. / 3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes. In: Biotechnology and Bioengineering. 2018 ; Vol. 115, No. 3. pp. 630-644.
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Correia, C, Koshkin, A, Duarte, P, Hu, D, Carido, M, Sebastião, MJ, Gomes-Alves, P, Elliott, DA, Domian, IJ, Teixeira, AP, Alves, PM & Serra, M 2018, '3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes', Biotechnology and Bioengineering, vol. 115, no. 3, pp. 630-644. https://doi.org/10.1002/bit.26504

3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes. / Correia, Cláudia; Koshkin, Alexey; Duarte, Patrícia; Hu, Dongjian; Carido, Madalena; Sebastião, Maria J.; Gomes-Alves, Patrícia; Elliott, David A.; Domian, Ibrahim J.; Teixeira, Ana Palma; Alves, Paula M.; Serra, Margarida.

In: Biotechnology and Bioengineering, Vol. 115, No. 3, 01.03.2018, p. 630-644.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 3D aggregate culture improves metabolic maturation of human pluripotent stem cell derived cardiomyocytes

AU - Correia, Cláudia

AU - Koshkin, Alexey

AU - Duarte, Patrícia

AU - Hu, Dongjian

AU - Carido, Madalena

AU - Sebastião, Maria J.

AU - Gomes-Alves, Patrícia

AU - Elliott, David A.

AU - Domian, Ibrahim J.

AU - Teixeira, Ana Palma

AU - Alves, Paula M.

AU - Serra, Margarida

PY - 2018/3/1

Y1 - 2018/3/1

N2 - Three-dimensional (3D) cultures of human pluripotent stem cell derived cardiomyocytes (hPSC-CMs) hold great promise for drug discovery, providing a better approximation to the in vivo physiology over standard two-dimensional (2D) monolayer cultures. However, the transition of CM differentiation protocols from 2D to 3D cultures is not straightforward. In this work, we relied on the aggregation of hPSC-derived cardiac progenitors and their culture under agitated conditions to generate highly pure cardiomyocyte aggregates. Whole-transcriptome analysis and 13C-metabolic flux analysis allowed to demonstrate at both molecular and fluxome levels that such 3D culture environment enhances metabolic maturation of hiPSC-CMs. When compared to 2D, 3D cultures of hiPSC-CMs displayed down-regulation of genes involved in glycolysis and lipid biosynthesis and increased expression of genes involved in OXPHOS. Accordingly, 3D cultures of hiPSC-CMs had lower fluxes through glycolysis and fatty acid synthesis and increased TCA-cycle activity. Importantly, we demonstrated that the 3D culture environment reproducibly improved both CM purity and metabolic maturation across different hPSC lines, thereby providing a robust strategy to derive enriched hPSC-CMs with metabolic features closer to that of adult CMs.

AB - Three-dimensional (3D) cultures of human pluripotent stem cell derived cardiomyocytes (hPSC-CMs) hold great promise for drug discovery, providing a better approximation to the in vivo physiology over standard two-dimensional (2D) monolayer cultures. However, the transition of CM differentiation protocols from 2D to 3D cultures is not straightforward. In this work, we relied on the aggregation of hPSC-derived cardiac progenitors and their culture under agitated conditions to generate highly pure cardiomyocyte aggregates. Whole-transcriptome analysis and 13C-metabolic flux analysis allowed to demonstrate at both molecular and fluxome levels that such 3D culture environment enhances metabolic maturation of hiPSC-CMs. When compared to 2D, 3D cultures of hiPSC-CMs displayed down-regulation of genes involved in glycolysis and lipid biosynthesis and increased expression of genes involved in OXPHOS. Accordingly, 3D cultures of hiPSC-CMs had lower fluxes through glycolysis and fatty acid synthesis and increased TCA-cycle activity. Importantly, we demonstrated that the 3D culture environment reproducibly improved both CM purity and metabolic maturation across different hPSC lines, thereby providing a robust strategy to derive enriched hPSC-CMs with metabolic features closer to that of adult CMs.

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