Perfusion decellularization of human and porcine lungs: bringing the matrix to clinical scale

The Journal of Heart and Lung Transplantation : the Official Publication of the International Society for Heart Transplantation
Sarah E GilpinHarald C Ott

Abstract

Organ engineering is a theoretical alternative to allotransplantation for end-stage organ failure. Whole-organ scaffolds can be created by detergent perfusion via the native vasculature, generating an acellular matrix suitable for recellularization with selected cell types. We aimed to up-scale this process, generating biocompatible scaffolds of a clinically relevant scale. Rat, porcine, and human lungs were decellularized by detergent perfusion at constant pressures. Collagen, elastin, and glycosaminoglycan content of scaffolds were quantified by colorimetric assays. Proteomic analysis was performed by microcapillary liquid chromatography tandem mass spectrometry. Extracellular matrix (ECM) slices were cultured with human umbilical vein endothelial cells (HUVEC), small airway epithelial cells (SAEC), or pulmonary alveolar epithelial cells (PAECs) and evaluated by time-lapse live cell microscopy and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Whole-organ culture was maintained under constant-pressure media perfusion after seeding with PAECs. Rat lungs were decellularized using: (1) sodium dodecyl sulfate (SDS), (2) sodium deoxycholate (SDC), or (3) 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesu...Continue Reading

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Citations

Mar 10, 2015·Cellular and Molecular Bioengineering·Darcy E WagnerDaniel J Weiss
May 31, 2014·Nature Protocols·Jacques P GuyetteHarald C Ott
Dec 3, 2014·The Canadian Journal of Cardiology·William S FosterDuncan J Stewart
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Dec 21, 2018·International Journal of Molecular Sciences·Andrea PorzionatoRaffaele De Caro

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