Advertisement

pp 1-21 | Cite as

Generation and Analysis of Pluripotent Stem Cell-Derived Cardiomyocytes and Endothelial Cells for High Content Screening Purposes

  • Tünde Berecz
  • Mária Husvéth-Tóth
  • Maxime Mioulane
  • Béla Merkely
  • Ágota Apáti
  • Gábor FöldesEmail author
Protocol
Part of the Methods in Molecular Biology book series

Abstract

Human-induced pluripotent stem cells (hiPSCs) and their differentiated derivatives became a new, promising source for in vitro screening techniques. Cell lines derived from healthy individuals can be applied for drug safety testing, while patient-derived cells provide a platform to model diseases in vitro and can be used as a tool for personalized medicine including specific drug efficacy testing and identification of new pharmacological targets as well as for tailoring pharmacological therapies. Efficient differentiation protocols yielding cardiomyocytes or endothelial cells derived from iPSCs have been developed recently. Phenotypic characterization and gene expression profiling of these derivatives can reveal clues for developmental and pathological questions. Moreover, functional analysis and cell-based assays using automated fluorescence imaging platform and high content analysis characterize cell type-specific profiles of hiPSC-derived cardiomyocytes (hiPSC-CM) and endothelial cells (hiPSC-EC) at the cellular and subcellular levels. This can be utilized in a platform which can provide multiple endpoint profiles of candidate compounds.

Keywords

Human-induced pluripotent stem cell Differentiation Cardiomyocyte Endothelial cell Toxicity Cell death Proliferation Automated high content imaging 

Notes

Acknowledgments

Supported by Medical Research Council (MR/R025002/1), BHF Centre of Regenerative Medicine, and the Hungarian National Research, Development and Innovation Fund (NVKP_16-1-2016-0017, NKFI-6 K128444 and OTKA K 128369).

References

  1. 1.
    Prasain N, Lee MR, Vemula S et al (2014) Differentiation of human pluripotent stem cells to cells similar to cord-blood endothelial colony-forming cells. Nat Biotechnol 32(11):1151–1157Google Scholar
  2. 2.
    Burridge PW, Matsa E, Shukla P et al (2014) Chemically defined generation of human cardiomyocytes. Nat Methods 11:855–860Google Scholar
  3. 3.
    Matsa E, Denning C (2012) In vitro uses of human pluripotent stem cell-derived cardiomyocytes. J Cardiovasc Transl Res 5:581–592Google Scholar
  4. 4.
    Mioulane M, Foldes G, Ali NN et al (2012) Quantification of cell death mechanisms in human pluripotent stem cell-derived cardiomyocytes. J Cardiovasc Transl Res 5:593–604Google Scholar
  5. 5.
    Hattori F, Chen H, Yamashita H et al (2010) Nongenetic method for purifying stem cell-derived cardiomyocytes. Nat Methods 7:61–66Google Scholar
  6. 6.
    Foldes G, Mioulane M, Wright JS et al (2011) Modulation of human embryonic stem cell-derived cardiomyocyte growth: a testbed for studying human cardiac hypertrophy? J Mol Cell Cardiol 50:367–376Google Scholar

Copyright information

© Springer Science+Business Media New York 2019

Authors and Affiliations

  • Tünde Berecz
    • 1
    • 2
  • Mária Husvéth-Tóth
    • 1
  • Maxime Mioulane
    • 3
  • Béla Merkely
    • 1
  • Ágota Apáti
    • 2
  • Gábor Földes
    • 1
    • 4
    Email author
  1. 1.Heart and Vascular CenterSemmelweis UniversityBudapestHungary
  2. 2.Institute of Enzymology, Research Centre for Natural SciencesBudapestHungary
  3. 3.Biosciences, Life Science Solutions, Thermo Fisher ScientificLondonUK
  4. 4.National Heart and Lung Institute, Imperial College London, Imperial Centre for Experimental and Translational MedicineLondonUK

Personalised recommendations