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Cardiac decellularisation with long-term storage and repopulation with canine peripheral blood progenitor cells

Authors

  • Bredon Crawford,

    1. Department of Chemical Engineering University of Waterloo 200 University Avenue West Waterloo, Ontario, Canada, N2L 3G1
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  • Sandeep Tharian Koshy,

    1. Harvard School of Engineering and Applied Sciences, 29 Oxford Street, Cambridge, Massachusetts 02138, U.S.A.
    2. Harvard-MIT Division of Health Sciences and Technology, Cambridge, Massachusetts, U.S.A.
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  • Geeta Jhamb,

    1. Department of Chemical Engineering University of Waterloo 200 University Avenue West Waterloo, Ontario, Canada, N2L 3G1
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  • Curtis Woodford,

    1. Institute of Biomaterials & Biomedical Engineering, Rosebrugh Building, 164 College Street, Room 407, University of Toronto, Toronto, Ontario, Canada, M5S 3G9
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  • Chistine M. Thompson,

    1. Department of Chemical Engineering University of Waterloo 200 University Avenue West Waterloo, Ontario, Canada, N2L 3G1
    2. Ontario Cancer Institute, Princess Margaret Hospital, 610 University Avenue, Toronto, Ontario, Canada, M5G 2M9
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  • Andrew S. Levy,

    1. Integrative Vascular Biology Laboratory, Department of Kinesiology, Faculty of Applied Health Sciences, University of Waterloo, 200 University Ave, West Waterloo, Ontario, Canada, N2L 3G1
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  • Jim W. E. Rush,

    1. Integrative Vascular Biology Laboratory, Department of Kinesiology, Faculty of Applied Health Sciences, University of Waterloo, 200 University Ave, West Waterloo, Ontario, Canada, N2L 3G1
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  • J. Guy Guillemette,

    1. Department of Chemistry, University of Waterloo, 200 University Avenue West, Waterloo, Ontario, Canada, N2L 3G1
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  • David Lillicrap,

    1. Department of Pathology & Molecular Medicine, Richardson Laboratory, Queen's University, 88 Stuart Street, Kingston, Ontario, Canada, K7L 3N6
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  • Eric Jervis

    Corresponding author
    1. Department of Chemical Engineering University of Waterloo 200 University Avenue West Waterloo, Ontario, Canada, N2L 3G1
    • Department of Chemical Engineering University of Waterloo 200 University Avenue West Waterloo, Ontario, Canada, N2L 3G1.
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  • Animals were sacrificed in the Integrative Vascular Biology Laboratory at the University of Waterloo. cBOECs were isolated in the Richardson Laboratory at Queen's University. Cell culture, modelling and reactor design and operation were performed in the Chemical Engineering Department at the University of Waterloo.

  • Disclosure statement: No competing financial interests exist.

Abstract

Perfusion decellularisation is a promising technique in scaffold production for tissue engineering. The similarity of decellularised extracellular matrix to native matrix is attractive for the development of compatible and functional constructs for therapeutic applications. Here, whole organs were processed for tissue culture with an emphasis on production staging. Adult rat hearts were perfusion-decellularised on a modified chromatography system that facilitated the study of flow properties and effluent material to better understand the transport processes of decellularisation. Tracer pulse experiments confirmed gross integrity of the ECM, and suggested the accumulation of debris in tissue lumen at late decellularisation. The rate of protein loss matched visual indications of decellularisation, slowing to zero as the tissue achieved a uniform appearance. Decellularised hearts were cryopreserved for up to 1 year before reconditioning including disinfection and soluble protein perfusion to remove adsorbed detergents. Canine blood outgrowth endothelial cells were seeded and cultured in a novel recellularisation apparatus with bubble-free aeration. After 9 days of culture, cells seeded in the lumen remained viable and exhibited well-attached morphology. This work presents approaches for the process analysis and non-destructive evaluation of decellularising tissue and demonstrates that cells may be successfully cultured on whole decellularised organs following long-term storage. © 2011 Canadian Society for Chemical Engineering

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