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Cardiac tissue engineering using perfusion bioreactor systems

Nature Protocols volume 3, pages 719–738 (2008)Cite this article

Abstract

This protocol describes tissue engineering of synchronously contractile cardiac constructs by culturing cardiac cell populations on porous scaffolds (in some cases with an array of channels) and bioreactors with perfusion of culture medium (in some cases supplemented with an oxygen carrier). The overall approach is 'biomimetic' in nature as it tends to provide in vivo-like oxygen supply to cultured cells and thereby overcome inherent limitations of diffusional transport in conventional culture systems. In order to mimic the capillary network, cells are cultured on channeled elastomer scaffolds that are perfused with culture medium that can contain oxygen carriers. The overall protocol takes 2–4 weeks, including assembly of the perfusion systems, preparation of scaffolds, cell seeding and cultivation, and on-line and end-point assessment methods. This model is well suited for a wide range of cardiac tissue engineering applications, including the use of human stem cells, and high-fidelity models for biological research.

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Figure 1: Cultivation of cardiac tissue constructs.
Figure 2: Predictions of oxygen profiles (micromolar) in a channel and tissue space of a construct with cell density of 1 × 108 cell ml−1, 100-μm channel diameter and 100-μm wall-to-wall spacing.
Figure 3: Generalized protocol flowchart.
Figure 4: A perfusion loop for cultivation of cardiac tissue constructs.
Figure 5: Scaffold fabrication.
Figure 7: Tissue culture in a generalized perfusion loop.
Figure 6: Seeding methods.

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Acknowledgements

The authors gratefully acknowledge research support of the work described in this protocol by the National Institutes of Health (R01 HL076485 and P41-EB002520 to G.V.-N.), National Science and Engineering Council (Discovery Grant to M.R.) and Canada Foundation for Innovation (Leaders Opportunity Fund to M.R.). The authors also thank Melissa A.N. Brown for help with taking pictures for Figures 4 and 7.

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Authors and Affiliations

  1. Department of Chemical Engineering and Applied Chemistry, Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada

    Milica Radisic

  2. Department of Biomedical Engineering, Columbia University, New York, New York, USA

    Anna Marsano, Robert Maidhof & Gordana Vunjak-Novakovic

  3. Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, USA

    Yadong Wang

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  1. Milica Radisic
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Correspondence to Gordana Vunjak-Novakovic.

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Radisic, M., Marsano, A., Maidhof, R. et al. Cardiac tissue engineering using perfusion bioreactor systems. Nat Protoc 3, 719–738 (2008). https://doi.org/10.1038/nprot.2008.40

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