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FRESH 3D Bioprinting a Ventricle-like Cardiac Construct Using Human Stem Cell-Derived Cardiomyocytes

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Cardiac Tissue Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2485))

Abstract

Here we describe a method to engineer a contractile ventricle-like chamber composed of human stem cell-derived cardiomyocytes using freeform reversible embedding of suspended hydrogels (FRESH) 3D bioprinting. To do this, we print a support structure using a collagen type I ink and a cellular component using a high-density cell ink supplemented with fibrinogen. The gelation of the collagen and the fibrinogen into fibrin is initiated by pH change and enzymatic crosslinking, respectively. Fabrication of the ventricle-like chamber is completed in three distinct phases: (i) materials preparation, (ii) bioprinting, and (iii) tissue maturation. In this protocol, we describe the method to print the construct from a high-density cell ink composed of human stem cell-derived cardiomyocytes and primary fibroblasts (~300 × 106 cells/mL) using our open-source dual-extruder bioprinter. Additional details are provided on FRESH support preparation, bioink preparation, dual-extruder needle alignment, print parameter selection, and post-processing. This protocol can also be adapted by altering the 3D model design, cell concentration, or cell type to FRESH 3D bioprint other cardiac tissue constructs.

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Author information

Authors and Affiliations

  1. Department of Materials Science and Engineering, Carnegie Mellon University, Pittsburgh, PA, USA

    Brian D. Coffin & Adam W. Feinberg

  2. Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA

    Andrew R. Hudson, Andrew Lee & Adam W. Feinberg

Authors
  1. Brian D. Coffin
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  2. Andrew R. Hudson
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  3. Andrew Lee
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  4. Adam W. Feinberg
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Corresponding author

Correspondence to Adam W. Feinberg .

Editor information

Editors and Affiliations

  1. School of Engineering, Brown University, Providence, RI, USA

    Kareen L.K. Coulombe

  2. Department of Biomedical Engineering, Tufts University, Medford, MA, USA

    Lauren D. Black III

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Coffin, B.D., Hudson, A.R., Lee, A., Feinberg, A.W. (2022). FRESH 3D Bioprinting a Ventricle-like Cardiac Construct Using Human Stem Cell-Derived Cardiomyocytes. In: Coulombe, K.L., Black III, L.D. (eds) Cardiac Tissue Engineering. Methods in Molecular Biology, vol 2485. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2261-2_5

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  • DOI: https://doi.org/10.1007/978-1-0716-2261-2_5

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2260-5

  • Online ISBN: 978-1-0716-2261-2

  • eBook Packages: Springer Protocols

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