title: Perfusion decellularization of human and porcine lungs: bringing the matrix to clinical scale.
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.
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School.
authors.forename: Sarah Elizabeth
authors.initials: SE
authors.lastname: Gilpin
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School.
authors.forename: Jacques P
authors.initials: JP
authors.lastname: Guyette
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Gonzalez
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School.
authors.forename: Xi
authors.initials: X
authors.lastname: Ren
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School.
authors.forename: John M
authors.initials: JM
authors.lastname: Asara
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Mathisen
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Division of Pediatric Surgery.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Vacanti
authors.affiliation: Center for Regenerative Medicine, Massachusetts General Hospital; Harvard Medical School; Division of Thoracic Surgery, Department of Surgery, Massachusetts General Hospital; Harvard Stem Cell Institute, Boston, Massachusetts. Electronic address: hott@partners.org.
authors.forename: Harald C
authors.initials: HC
authors.lastname: Ott
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D057005:Bioengineering / Q000379:methods*; D002478:Cells, Cultured; D002793:Cholic Acids / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000494:pharmacology*; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology; D005109:Extracellular Matrix / Q000187:drug effects*; D006801:Humans; D008168:Lung / Q000166:cytology / Q000187:drug effects*; D023421:Models, Animal; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D013552:Swine; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D002793:Cholic Acids; D003902:Detergents; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate
keywords: acellular matrix; decellularization; organ engineering; organ scaffolds; recellularization