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      Re-epithelialization of whole porcine kidneys with renal epithelial cells

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          Abstract

          Decellularized porcine kidneys were recellularized with renal epithelial cells by three methods: perfusion through the vasculature under high pressure, perfusion through the ureter under high pressure, or perfusion through the ureter under moderate vacuum. Histology, scanning electron microscopy, confocal microscopy, and magnetic resonance imaging were used to assess vasculature preservation and the distribution of cells throughout the kidneys. Cells were detected in the magnetic resonance imaging by labeling them with iron oxide. Perfusion of cells through the ureter under moderate vacuum (40 mmHg) produced the most uniform distribution of cells throughout the kidneys.

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          Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds.

          The definitive treatment for end-stage organ failure is orthotopic transplantation. However, the demand for transplantation far exceeds the number of available donor organs. A promising tissue-engineering/regenerative-medicine approach for functional organ replacement has emerged in recent years. Decellularization of donor organs such as heart, liver, and lung can provide an acellular, naturally occurring three-dimensional biologic scaffold material that can then be seeded with selected cell populations. Preliminary studies in animal models have provided encouraging results for the proof of concept. However, significant challenges for three-dimensional organ engineering approach remain. This manuscript describes the fundamental concepts of whole-organ engineering, including characterization of the extracellular matrix as a scaffold, methods for decellularization of vascular organs, potential cells to reseed such a scaffold, techniques for the recellularization process and important aspects regarding bioreactor design to support this approach. Critical challenges and future directions are also discussed.
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            Regeneration and orthotopic transplantation of a bioartificial lung.

            About 2,000 patients now await a donor lung in the United States. Worldwide, 50 million individuals are living with end-stage lung disease. Creation of a bioartificial lung requires engineering of viable lung architecture enabling ventilation, perfusion and gas exchange. We decellularized lungs by detergent perfusion and yielded scaffolds with acellular vasculature, airways and alveoli. To regenerate gas exchange tissue, we seeded scaffolds with epithelial and endothelial cells. To establish function, we perfused and ventilated cell-seeded constructs in a bioreactor simulating the physiologic environment of developing lung. By day 5, constructs could be perfused with blood and ventilated using physiologic pressures, and they generated gas exchange comparable to that of isolated native lungs. To show in vivo function, we transplanted regenerated lungs into orthotopic position. After transplantation, constructs were perfused by the recipient's circulation and ventilated by means of the recipient's airway and respiratory muscles, and they provided gas exchange in vivo for up to 6 h after extubation.
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              Regeneration and Experimental Orthotopic Transplantation of a Bioengineered Kidney

              Over 100,000 individuals in the United States currently await kidney transplantation, while 400,000 individuals live with end-stage kidney disease requiring hemodialysis. The creation of a transplantable graft to permanently replace kidney function would address donor organ shortage and the morbidity associated with immunosuppression. Such a bioengineered graft must have the kidney’s architecture and function, and permit perfusion, filtration, secretion, absorption, and drainage of urine. We decellularized rat, porcine, and human kidneys by detergent perfusion, yielding acellular scaffolds with vascular, cortical and medullary architecture, collecting system and ureters. To regenerate functional tissue, we seeded rat kidney scaffolds with epithelial and endothelial cells, then perfused these cell-seeded constructs in a whole organ bioreactor. The resulting grafts produced rudimentary urine in vitro when perfused via their intrinsic vascular bed. When transplanted in orthotopic position in rat, the grafts were perfused by the recipient’s circulation, and produced urine via the ureteral conduit in vivo.
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                Author and article information

                Journal
                J Tissue Eng
                J Tissue Eng
                TEJ
                sptej
                Journal of Tissue Engineering
                SAGE Publications (Sage UK: London, England )
                2041-7314
                03 July 2017
                Jan-Dec 2017
                : 8
                : 2041731417718809
                Affiliations
                [1 ]Department of Chemical Engineering, Brigham Young University, Provo, UT, USA
                [2 ]Department of Genetics and Biotechnology, Brigham Young University, Provo, UT, USA
                [3 ]Department of Electrical Engineering, Brigham Young University, Provo, UT, USA
                [4 ]Department of Physiology and Developmental Biology, Brigham Young University, Provo, UT, USA
                [5 ]Department of Biology, Brigham Young University, Provo, UT, USA
                Author notes
                [*]Alonzo Cook, Department of Chemical Engineering, Brigham Young University, Provo, UT 84602, USA. Email: cook@ 123456byu.edu
                Article
                10.1177_2041731417718809
                10.1177/2041731417718809
                5513523
                3a1844e2-4e0a-4f2c-895f-08d8930574af
                © The Author(s) 2017

                This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License ( http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page ( http://www.uk.sagepub.com/aboutus/openaccess.htm).

                History
                : 8 May 2017
                : 13 June 2017
                Categories
                Original Article
                Custom metadata
                January-December 2017

                Biomedical engineering
                recellularization,kidney,epithelial cells,organ regeneration,magnetic resonance imaging

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