title: Rapid porcine lung decellularization using a novel organ regenerative control acquisition bioreactor.
abstract: To regenerate discarded lungs that would not normally be used for transplant, ex vivo reseeding after decellularization may produce organs suitable for clinical transplantation and therefore close the donor gap. Organ regenerative control acquisition (Harvard Biosciences, Holliston, MA), a novel bioreactor system that simulates physiological conditions, was used to evaluate a method of rapid decellularization. Although most current decellularization methods are 24-72 hours, we hypothesized that perfusing porcine lungs with detergents at higher pressures for less time would yield comparable bioscaffolds suitable for future experimentation. Methods involved perfusion of 1% Triton X-100 (Triton) and 0.1% sodium dodecyl sulfate at varied physiological flow rates. Architecture of native and decellularized lungs was analyzed with hematoxylin and eosin (H&E) staining, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). Dry gas and liquid ventilation techniques were introduced. Our 7 hour decellularization procedure removes nuclear material while maintaining architecture. Bioscaffolds have the microarchitecture for reseeding of stem cells. Hematoxylin and eosin staining suggested removal of nuclear material, whereas SEM and TEM imaging demonstrated total removal of cells with structural architecture preserved. This process can lead to clinical implementation, thereby increasing the availability of human lungs for transplantation.
authors.affiliation: From the *Division of Cardiothoracic Surgery, Department of Surgery, University of Arizona College of Medicine, Tucson, Arizona; †University of Arizona Medical Center, Tucson, Arizona; ‡Department of Physiology, University of Arizona College of Medicine, Tucson, Arizona; §Department of Biomedical Engineering, University of Arizona College of Medicine, Tucson, Arizona; and ¶University of Arizona College of Medicine, Tucson, Arizona.
authors.forename: Zain
authors.initials: Z
authors.lastname: Khalpey
authors.forename: Ning
authors.initials: N
authors.lastname: Qu
authors.forename: Courtney
authors.initials: C
authors.lastname: Hemphill
authors.forename: Anthony V
authors.initials: AV
authors.lastname: Louis
authors.forename: Alice S
authors.initials: AS
authors.lastname: Ferng
authors.forename: Tiffany G
authors.initials: TG
authors.lastname: Son
authors.forename: Katherine
authors.initials: K
authors.lastname: Stavoe
authors.forename: Kitsie
authors.initials: K
authors.lastname: Penick
authors.forename: Phat L
authors.initials: PL
authors.lastname: Tran
authors.forename: John
authors.initials: J
authors.lastname: Konhilas
authors.forename: Destiny S
authors.initials: DS
authors.lastname: Lagrand
authors.forename: Joe G N
authors.initials: JG
authors.lastname: Garcia
mesh_terms: D000818:Animals; D021522:Bioartificial Organs; D019149:Bioreactors; D004247:DNA / Q000032:analysis; D006801:Humans; D008168:Lung / Q000166:cytology* / Q000502:physiology; D016040:Lung Transplantation; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D012038:Regeneration / Q000502:physiology; D034421:Sus scrofa; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: D004247:DNA