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Developing HiPSC Derived Serum Free Embryoid Bodies for the Interrogation of 3-D Stem Cell Cultures Using Physiologically Relevant Assays
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A complete workflow for the differentiation and the dissociation of hiPSC-derived cardiospheres

Benjamin Fischer1, Anna Meier1, Annika Dehne1

  • 1Fraunhofer Institute for Biomedical Engineering, Joseph-von-Fraunhofer-Weg 1, Sulzbach 66280, Germany.

Stem Cell Research
|September 16, 2018
PubMed

Insights

This study presents a new workflow for producing human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) using a dynamic suspension bioreactor. This method improves yield and viability for cardiovascular research and drug discovery.

Area of Science:

  • Cardiovascular Research
  • Stem Cell Biology
  • Biotechnology

Background:

  • Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are crucial for cardiovascular research, disease modeling, and drug discovery.
  • Current methods for hiPSC-CM production face limitations in yield and efficiency, hindering their therapeutic and research potential.

Purpose of the Study:

  • To develop a complete workflow for producing ready-to-use hiPSC-CMs in a dynamic suspension bioreactor.
  • To improve the efficiency, yield, and viability of hiPSC-CM production compared to existing methods.

Main Methods:

  • Utilized a dynamic suspension bioreactor for efficient and reproducible differentiation of hiPSCs into cardiospheres.
  • Developed a novel papain-based dissociation method for hiPSC-CMs, offering higher yield and viability than TrypLE and Accutase.
  • Performed molecular and functional analyses to confirm the identity and functionality of dissociated hiPSC-CMs.

Main Results:

  • The dynamic suspension bioreactor method resulted in enhanced physiological maturation of cardiospheres compared to static 3D induction.
  • The papain-based dissociation method significantly increased hiPSC-CM yield and viability.
  • hiPSC-CMs retained their identity and functionality after dissociation and reseeding for downstream applications.

Conclusions:

  • The presented workflow provides an efficient and scalable method for producing high-quality hiPSC-CMs.
  • This optimized production of hiPSC-CMs can overcome current bottlenecks, advancing cardiovascular research, disease modeling, and drug discovery.
  • The ready-to-use hiPSC-CMs are suitable for various downstream applications, including monolayer and spheroid cultures.

Keywords:
3D bioreactorCardiac inductionCardiomyocytesPapain dissociationhiPSCs

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