Cited by 263Tissue Engineering and Regenerative Medicine3D Printing in Biomedical ResearchTransplantation: Methods and Outcomes

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

Gilpin, Sarah Elizabeth, Guyette, Jacques P., Gonzalez, Gabriel, Ren, Xi, Asara, John M., Mathisen, Douglas J., Vacanti, Joseph P., Ott, Harald C.·2014·The Journal of Heart and Lung Transplantation

DOI 10.1016/j.healun.2013.10.030

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Abstract

Background 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.Methods 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.Results Rat lungs were decellularized using: (1) sodium dodecyl sulfate (SDS), (2) sodium deoxycholate (SDC), or (3) 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). Resulting scaffolds showed comparable loss of DNA but greatest preservation of ECM components in SDS-decellularized lungs. Porcine (n = 10) and human (n = 7) lungs required increased SDS concentration, perfusion pressures, and time to achieve decellularization as determined by loss of DNA, with preservation of intact matrix composition and lung architecture. Proteomic analysis of human decellularized lungs further confirmed ECM preservation. Recellularization experiments confirmed scaffold biocompatibility when cultured with mature cell phenotypes and scaffold integrity for the duration of biomimetic culture.Conclusions SDS-based perfusion decellularization can be applied to whole porcine and human lungs to generate biocompatible organ scaffolds with preserved ECM composition and architecture.

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Authors: Gilpin, S. E., Guyette, J. P., Gonzalez, G., Ren, X., Asara, J. M., Mathisen, D. J., Vacanti, J. P., & Ott, H. C.
Affiliations: Harvard University (US)
Massachusetts General Hospital (US)
Corresponding author: Ott, Harald C.
ORCID: Gilpin, Sarah Elizabeth
Ren, Xi
Asara, John M.
Mathisen, Douglas J.
Ott, Harald C.
Venue: The Journal of Heart and Lung Transplantation
Publisher: Elsevier BV
Published: 2014-03
Volume: 33
Issue: 3
Pages: 298-308
ISSN: 1053-2498 (ISSN-L)
DOI: 10.1016/j.healun.2013.10.030
OpenAlex: W2047611832
Semantic Scholar: 253777fdc37ab99c976b2319df807c9150ac0b39
Cited by: 263
References: 41
Citations by year: 2026: 10 · 2025: 6 · 2024: 12 · 2023: 18 · 2022: 17 · 2021: 33 · 2020: 25 · 2019: 24 · 2018: 29 · 2017: 26 · 2016: 21 · 2015: 27 · 2014: 14
FWCI: 20.37
Type: journal article
Language: en
Open access: closed
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
Locations: The Journal of Heart and Lung Transplantation (publishedVersion)
PubMed (publishedVersion)
Funders: National Institutes of Health (1dp2od008749-01)
Topics: Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
Health Sciences › Medicine › Surgery › Transplantation: Methods and Outcomes
Keywords: perfusion decellularization, lung scaffolds, extracellular matrix preservation, sodium dodecyl sulfate, sodium deoxycholate, CHAPS, human lung, porcine lung, recellularization, alveolar epithelial cells, endothelial cells, small airway epithelial cells, whole organ culture, organ engineering, scaffold biocompatibility
MeSH terms: Animals, Biocompatible Materials, Cells, Cultured, Cholic Acids, Deoxycholic Acid, Detergents, Dose-Response Relationship, Drug, Endothelium, Vascular, Epithelial Cells, Extracellular Matrix, Humans, Lung, Perfusion, Sodium Dodecyl Sulfate, Swine, Rats, Sprague-Dawley, Models, Animal, Rats, Tissue Scaffolds, Bioengineering

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

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