Stem Cell-Derived Cardiac Spheroids as 3D In Vitro Models of the Human Heart Microenvironment

  • Madeline Campbell
  • Mamta Chabria
  • Gemma A. Figtree
  • Liudmila Polonchuk
  • Carmine GentileEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2002)


Our laboratory has recently developed a novel three-dimensional in vitro model of the human heart, which we call the vascularized cardiac spheroid (VCS). These better recapitulate the human heart’s cellular and extracellular microenvironment compared to the existing in vitro models. To achieve this, human-induced pluripotent stem cell (iPSC)-derived cardiomyocytes, cardiac fibroblasts, and human coronary artery endothelial cells are co-cultured in hanging drop culture in ratios similar to those found in the human heart in vivo. The resulting three-dimensional cellular organization, extracellular matrix, and microvascular network formation throughout the VCS has been shown to mimic the one present in the human heart tissue. Therefore, VCSs offer a promising platform to study cardiac physiology, disease, and pharmacology, as well as bioengineering constructs to regenerate heart tissue.


3D cultures Bioprinting Cardiovascular regeneration Heart microenvironment Induced pluripotent stem cells Niche Tissue bioengineering Vascularized cardiac spheroid 



This study was supported by a Postdoctoral Marcus Blackmore Fellowship from the Heart Research Institute and a Kick-Start Grant, a Cardiothoracic Surgery Research Grant Scheme and a CDIP Industry & Community Engagement Fund 2017 from the University of Sydney to CG, by an NHMRC Project Grant (APP1129685) to GF and CG, and by a Roche Post-doctoral Fellowship to MC. We would like to thank Dr. John Russell Brereton (Royal North Shore Hospital, Sydney) for his support, Dr. Christine Chuang (University of Copenhagen) for help with the ECM studies, and Dr. Louise Cole (University of Sydney) for their assistance with confocal imaging.


  1. 1.
    Gentile C (2016) Filling the gaps between the in vivo and in vitro microenvironment: engineering of spheroids for stem cell technology. Curr Stem Cell Res Ther 11:652–665CrossRefGoogle Scholar
  2. 2.
    Fennema E, Rivron N, Rouwkema J et al (2013) Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol 31:108–115. CrossRefPubMedGoogle Scholar
  3. 3.
    Hakkinen KM, Harunaga JS, Doyle AD, Yamada KM (2011) Direct comparisons of the morphology, migration, cell adhesions, and actin cytoskeleton of fibroblasts in four different three-dimensional extracellular matrices. Tissue Eng A 17:713–724. CrossRefGoogle Scholar
  4. 4.
    Edmondson R, Broglie JJ, Adcock AF, Yang L (2014) Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev Technol 12:207–218. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Zuppinger C (2016) 3D culture for cardiac cells. Mol Cell Res 1863:1873–1881. CrossRefGoogle Scholar
  6. 6.
    Baker BM, Chen CS (2012) Deconstructing the third dimension—how 3D culture microenvironments alter cellular cues. J Cell Sci 125:3015–3024CrossRefGoogle Scholar
  7. 7.
    Polonchuk L, Chabria M, Badi L et al (2017) Cardiac spheroids as promising in vitro models to study the human heart microenvironment. Sci Rep 7:1–12. CrossRefGoogle Scholar
  8. 8.
    Figtree GA, Bubb KJ, Tang O et al (2017) Vascularized cardiac spheroids as novel 3D in vitro models to study cardiac fibrosis. Cells Tissues Organs 204:191–198. CrossRefPubMedGoogle Scholar
  9. 9.
    Benam KH, Dauth S, Hassell B et al (2015) Engineered in vitro disease models. Annu Rev Pathol 10:195–262. CrossRefPubMedGoogle Scholar
  10. 10.
    Fitzgerald KA, Malhotra M, Curtin CM et al (2015) Life in 3D is never flat: 3D models to optimise drug delivery. J Control Release 215:39–54. CrossRefPubMedGoogle Scholar
  11. 11.
    Visconti RP, Kasyanov V, Gentile C et al (2010) Towards organ printing: engineering an intra-organ branched vascular tree. Expert Opin Biol Ther 10:409–420. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Madeline Campbell
    • 1
  • Mamta Chabria
    • 2
  • Gemma A. Figtree
    • 1
  • Liudmila Polonchuk
    • 2
  • Carmine Gentile
    • 1
    • 3
    • 4
    Email author
  1. 1.Sydney Medical SchoolUniversity of SydneySydneyAustralia
  2. 2.Roche Pharma Research and Early Development, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd.BaselSwitzerland
  3. 3.Beth Israel Deaconess Medical CenterHarvard Medical SchoolBostonUSA
  4. 4.Kolling Institute, Royal North Shore HospitalSydneyAustralia

Personalised recommendations