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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