Combining Hypoxia and Bioreactor Hydrodynamics Boosts Induced Pluripotent Stem Cell Differentiation Towards Cardiomyocytes

Cláudia Correia, Margarida Serra, Nuno Espinha, Marcos Sousa, Ana Catarina Montes, Karsten Burkert, Yunjie Zheng, Jürgen Hescheler, Manuel J T Carrondo, Tomo Šarić, Paula M. Alves

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Cardiomyocytes (CMs) derived from induced pluripotent stem cells (iPSCs) hold great promise for patient-specific disease modeling, drug screening and cell therapy. However, existing protocols for CM differentiation of iPSCs besides being highly dependent on the application of expensive growth factors show low reproducibility and scalability. The aim of this work was to develop a robust and scalable strategy for mass production of iPSC-derived CMs by designing a bioreactor protocol that ensures a hypoxic and mechanical environment. Murine iPSCs were cultivated as aggregates in either stirred tank or WAVE bioreactors. The effect of dissolved oxygen and mechanical forces, promoted by different hydrodynamic environments, on CM differentiation was evaluated. Combining a hypoxia culture (4 % O2 tension) with an intermittent agitation profile in stirred tank bioreactors resulted in an improvement of about 1000-fold in CM yields when compared to normoxic (20 % O2 tension) and continuously agitated cultures. Additionally, we showed for the first time that wave-induced agitation enables the differentiation of iPSCs towards CMs at faster kinetics and with higher yields (60 CMs/input iPSC). In an 11-day differentiation protocol, clinically relevant numbers of CMs (2.3 × 109 CMs/1 L) were produced, and CMs exhibited typical cardiac sarcomeric structures, calcium transients, electrophysiological profiles and drug responsiveness. This work describes significant advances towards scalable cardiomyocyte differentiation of murine iPSC, paving the way for the implementation of this strategy for mass production of their human counterparts and their use for cardiac repair and cardiovascular research.

Original languageEnglish
Pages (from-to)786-801
Number of pages16
JournalStem Cell Reviews and Reports
Volume10
Issue number6
DOIs
Publication statusPublished - 2014

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Induced Pluripotent Stem Cells
Bioreactors
Hydrodynamics
Cardiac Myocytes
Cell Differentiation
Hypoxia
Preclinical Drug Evaluations
Cell- and Tissue-Based Therapy
Intercellular Signaling Peptides and Proteins

Keywords

  • Bioreactor hydrodynamics
  • Cardiomyocyte differentiation
  • Hypoxia
  • Induced pluripotent stem cells
  • Mass production
  • Mechanical environment

Cite this

Correia, Cláudia ; Serra, Margarida ; Espinha, Nuno ; Sousa, Marcos ; Montes, Ana Catarina ; Burkert, Karsten ; Zheng, Yunjie ; Hescheler, Jürgen ; Carrondo, Manuel J T ; Šarić, Tomo ; Alves, Paula M. / Combining Hypoxia and Bioreactor Hydrodynamics Boosts Induced Pluripotent Stem Cell Differentiation Towards Cardiomyocytes. In: Stem Cell Reviews and Reports. 2014 ; Vol. 10, No. 6. pp. 786-801.
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Combining Hypoxia and Bioreactor Hydrodynamics Boosts Induced Pluripotent Stem Cell Differentiation Towards Cardiomyocytes. / Correia, Cláudia; Serra, Margarida; Espinha, Nuno; Sousa, Marcos; Montes, Ana Catarina; Burkert, Karsten; Zheng, Yunjie; Hescheler, Jürgen; Carrondo, Manuel J T; Šarić, Tomo; Alves, Paula M.

In: Stem Cell Reviews and Reports, Vol. 10, No. 6, 2014, p. 786-801.

Research output: Contribution to journalArticle

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