Granted Patent
B2
US 12,337,078 · App. 18/170,669 · Granted Jun 24, 2025
Organ/tissue decellularization, framework maintenance and recellularization
Assignee:
WAKE FOREST UNIVERSITY HEALTH SCIENCES
A61L27/3633A61L27/3604A61L27/3687A61L27/3808A61L27/3834A61L27/54A61L2300/256A61L2300/42A61L2430/26A61L2430/28A61L2430/40
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Abstract
Methods for decellularizing organs and tissues in vitro and in vivo are provided, as are methods of maintaining organ and tissue frameworks and methods of recellularizing organs and tissues, thereby providing an approach to needed organs or tissues.
Claims (15)
1. A method of regenerating an organ from an acellular organ scaffold for a subject in need comprising:
(a) providing an acellular organ scaffold, wherein the organ is an ex vivo organ that is diseased, injured, damaged, or discarded for a medical reason, and wherein the organ is decellularized by a method comprising the steps of:
(i) perfusing the ex vivo organ with at least 50 volumes of a detergent, wherein the volume is the volume of the organ being perfused; and
(ii) rinsing the ex vivo organ with at least 50 volumes of a neutral buffer, wherein the perfusing and rinsing are performed using at least two different fluid inlets and at least one different fluid outlet, thereby resulting in a decellularized organ;
(b) contacting the acellular organ scaffold of step (a) with a cell characteristic of the acellular organ scaffold;
(c) incubating the acellular organ scaffold and cell ex vivo under ramping perfusion conditions to produce an incubated organ scaffold, wherein the perfusion comprises delivery of at least 1×10 8 cells at a rate spanning 2 ml/minute to 20 ml/minute; and
(d) implanting the incubated organ scaffold into the subject and establishing at least two vascular connections of the scaffold vasculature and the subject vasculature, thereby producing a regenerated organ from an acellular organ scaffold.
2. The method of claim 1 wherein the cell is obtained from the subject.
3. The method of claim 1 wherein the scaffold is contacted with at least 5×10 7 cells.
4. The method of claim 1 wherein the cell is a vascular endothelial cell.
5. The method of claim 1 further comprising attachment to the scaffold of an antibody product specifically binding to the cell.
6. The method of claim 5 wherein the antibody product comprises an antigen binding site for CD31.
7. The method of claim 1 further comprising attachment of an anti-thrombotic composition to the scaffold.
8. The method of claim 7 wherein the anti-thrombotic composition is heparin.
9. The method of claim 1 wherein the organ is a kidney, a pancreas or a liver.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 12, 2025
From: ORLANDO, GIUSEPPE; SOKER, SHAY
To: WAKE FOREST UNIVERSITY HEALTH SCIENCES
Reel/Frame 071397/0574 →
Continuity (3)
Provisional Application
61934429
· Jan 31, 2014
References Cited (174)
US 4946778A
· Ladner et al.
· 1990
[cited by applicant]
US 5225539A
· Winter
· 1993
[cited by applicant]
US 5403484A
· Ladner et al.
· 1995
[cited by applicant]
US 5545806A
· Lonberg et al.
· 1996
[cited by applicant]
US 5569825A
· Lonberg et al.
· 1996
[cited by applicant]
US 5571698A
· Ladner et al.
· 1996
[cited by applicant]
US 5585089A
· Queen et al.
· 1996
[cited by applicant]
US 5639641A
· Pedersen et al.
· 1997
[cited by applicant]
US 5693761A
· Queen et al.
· 1997
[cited by applicant]
US 5693762A
· Queen et al.
· 1997
[cited by applicant]
US 5702892A
· Mulligan-Kehoe
· 1997
[cited by applicant]
US 5714352A
· Jakobovits
· 1998
[cited by applicant]
US 5780279A
· Matthews et al.
· 1998
[cited by applicant]
US 5821047A
· Garrard et al.
· 1998
[cited by applicant]
US 5824520A
· Mulligan-Kehoe
· 1998
[cited by applicant]
US 5837500A
· Ladner et al.
· 1998
[cited by applicant]
US 5855885A
· Smith et al.
· 1999
[cited by applicant]
US 5858657A
· Winter et al.
· 1999
[cited by applicant]
US 5871907A
· Winter et al.
· 1999
[cited by applicant]
US 5969108A
· McCafferty et al.
· 1999
[cited by applicant]
US 6057098A
· Buechler et al.
· 2000
[cited by applicant]
US 6225447B1
· Winter et al.
· 2001
[cited by applicant]
US 6265150B1
· Terstappen et al.
· 2001
[cited by applicant]
US 6376244B1
· Atala
· 2002
[cited by examiner]
US 20020197266A1
· Debinski
· 2002
[cited by applicant]
US 20050282233A1
· Eriksson et al.
· 2005
[cited by applicant]
US 20090191608A1
· Matsumoto
· 2009
[cited by applicant]
EP 0239400B1
· 1994
[cited by applicant]
GB 2188638A
· 1987
[cited by applicant]
WO WO2005087812A1
· 2005
[cited by applicant]
WO WO2007025233A1
· 2007
[cited by applicant]
Badylak et al. Annu Rev Biomed Eng 13:27 (Year: 2011).
[cited by examiner]
Abbasoglu O, Liver transplantation: yesterday, today and tomorrow,
[cited by applicant]
Abt et al., Survival following liver transplantation from non-heart-beating donors,
[cited by applicant]
Amann et al., Therapeutic window of MuS110, a single-chain antibody construct bispecific for murine EpCAM and murine CD3,
[cited by applicant]
American Diabetes Association, Diagnosis and classification of diabetes mellitus,
[cited by applicant]
American Diabetes Association, Economic costs of diabetes in the U.S. in 2012,
[cited by applicant]
Arbiser et al., Oncogenic H-ras stimulates tumor angiogenesis by two distinct pathways,
[cited by applicant]
Arenas-Herrera et al., Decellularization for whole organ bioengineering,
[cited by applicant]
Arruebo et al., Antibody-conjugated nanoparticles for biomedical applications,
[cited by applicant]
Atala, Organ preservation, organ and cell transplantation, tissue engineering, and regenerative medicine: the terms may change, but the goals remain the same,
[cited by applicant]
Atala et al., Engineering complex tissues,
[cited by applicant]
Atala et al., Tissue-engineered autologous bladders for patients needing cystoplasty,
[cited by applicant]
Bach et al., Genetics of transplantation: the major histocompatibility complex,
[cited by applicant]
Badylak et al., Engineered whole organs and complex tissues,
[cited by applicant]
Badylak et al., Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds,
[cited by applicant]
Baiguera et al., Chorioallantoic membrane for in vivo investigation of tissue-engineered construct biocompatibility,
[cited by applicant]
Baptista et al., The use of whole organ decellularization for the generation of a vascularized liver organoid,
[cited by applicant]
Barakat et al., Use of decellularized porcine liver for engineering humanized liver organ,
[cited by applicant]
Bissell et al., How does the extracellular matrix direct gene expression?
[cited by applicant]
Bonandrini et al., Recellularization of well-preserved acellular kidney scaffold using embryonic stem cells,
[cited by applicant]
Boyle et al., Projection of the year 2050 burden of diabetes in the US adult population: dynamic modeling of incidence, mortality, and prediabetes prevalence,
[cited by applicant]
Brien et al., Protection by immunoglobulin dual-affinity retargeting antibodies against dengue virus,
[cited by applicant]
Brischwein et al., MT110: a novel bispecific single-chain antibody construct with high efficacy in eradicating established tumors,
[cited by applicant]
Casavilla et al., Experience with liver and kidney allografts from non-heart-beating donors,
[cited by applicant]
Chazan et al., The clinical spectrum of renal osteodystrophy in 57 chronic hemodialysis patients: a correlation between biochemical parameters and bone pathology findings,
[cited by applicant]
Choi et al., Development of a porcine renal extracellular matrix scaffold as a platform for kidney regeneration,
[cited by applicant]
Chung et al., Bladder reconstitution with bone marrow derived stem cells seeded on small intestinal submucosa improves morphological and molecular composition,
[cited by applicant]
Cioffi et al., EpCAM/CD3-Bispecific T-cell engaging antibody MT110 eliminates primary human pancreatic cancer stem cells,
[cited by applicant]
Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R Liss Inc., New York: 77-96 (1985).
[cited by applicant]
Cortiella et al., Influence of acellular natural lung matrix on murine embryonic stem cell differentiation and tissue formation,
[cited by applicant]
Cote et al., Generation of human monoclonal antibodies reactive with cellular antigens,
[cited by applicant]
Crapo et al., An overview of tissue and whole organ decellularization processes.
[cited by applicant]
Daghini et al., Assessment of renal hemodynamics and function in pigs with 64-section multidetector CT: comparison with electron-beam CT,
[cited by applicant]
Daley et al., Extracellular matrix dynamics in development and regenerative medicine,
[cited by applicant]
Donati et al., Extracorporeal detoxification for hepatic failure using molecular adsorbent recirculating system: depurative efficiency and clinical results in a longterm follow-up,
[cited by applicant]
Esmatjes et al., The utility of the C9peptide in the phenotyping of patient candidates for pancreas transplantation,
[cited by applicant]
Faulk et al., Role of the extracellular matrix in whole organ engineering,
[cited by applicant]
Fournier et al., Bispecific antibodies and trispecific immunocytokines for targeting the immune system against cancer: preparing for the future,
[cited by applicant]
Gattazzo et al., Extracellular matrix: a dynamic microenvironment for stem cell niche,
[cited by applicant]
Gilbert, Strategies for tissue and organ decellularization,
[cited by applicant]
Brody et al., Aptamers as therapeutic and diagnostic agents, J. Biotechnol. 74(1): 5-13 (2000).
[cited by applicant]
Gray, Anatomy of the human body: 2i. The liver, (20th ed.): 1-54 (1918).
[cited by applicant]
Groffen et al., Agrin is a major heparan sulfate proteoglycan in the human glomerular basement membrane,
[cited by applicant]
Hammel et al., Regression of liver fibrosis after biliary drainage in patients with chronic pancreatitis and stenosis of the common bile duct,
[cited by applicant]
Haskard et al., The production of human monoclonal autoantibodies from patients with rheumatoid arthritis by the EBV-hybridoma technique,
[cited by applicant]
Hata et al., Transplantation of engineered chimeric liver with autologous hepatocytes and xenobiotic scaffold,
[cited by applicant]
Hoerstrup et al., Functional growth in tissue-engineered living, vascular grafts: follow-up at 100 weeks in a large animal model,
[cited by applicant]
Huse et al., Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda,
[cited by applicant]
Hynes. The extracellular matrix: not just pretty fibrils,
[cited by applicant]
Iannotti et al., Porcine small intestine submucosa augmentation of surgical repair of chronic two-tendon rotator cuff tears. A randomized, controlled trial,
[cited by applicant]
Jain et al., Engineering vascularized tissue,
[cited by applicant]
Janeway et al. (eds.), Immunobiology, Garland Publishing, New York, (5th ed.): (2001).
[cited by applicant]
Johnson et al., Effector cell recruitment with novel Fv-based dual-affinity re-targeting protein leads to potent tumor cytolysis and in vivo B-cell depletion,
[cited by applicant]
Jones et al., Replacing the complementarity-determining regions in a human antibody with those from a mouse,
[cited by applicant]
Kabat et al., Sequences of proteins of immunological interest, U.S. Department of Health and Human Services, (1983).
[cited by applicant]
Kanasaki et al., Integrin beta1-mediated matrix assembly and signalling are critical for the normal development and function of the kidney glomerulus,
[cited by applicant]
Ko et al., Enhanced re-endothelialization of acellular kidney scaffolds for whole organ engineering via antibody conjugation of vasculatures,
[cited by applicant]
Ko et al., Targeting mesenchymal stem cells to activated endothelial cells,
[cited by applicant]
Kohler et al., Continuous cultures of fused cells secreting antibody of predefined specificity,
[cited by applicant]
Kortt et al., Dimeric and trimeric antibodies: high avidity scFvs for cancer targeting,
[cited by applicant]
Kozbor et al., The production of monoclonal antibodies from human lymphocytes,
[cited by applicant]
Lang et al., Three-dimensional culture of hepatocytes on porcine liver tissue-derived extracellular matrix,
[cited by applicant]
Lawton et al., Early stenosis of medtronic mosaic porcine valves in the aortic position,
[cited by applicant]
Lin et al., Spironolactone ameliorates podocytic adhesive capacity via restoring integrin alpha 3 expression in streptozotocin-induced diabetic rats,
[cited by applicant]
Macchiarini et al., Clinical transplantation of a tissue-engineered airway,
[cited by applicant]
Manelli et al., 3D analysis of SEM images of corrosion casting using adaptive stereo matching,
[cited by applicant]
Marcellin et al., Regression of cirrhosis during treatment with tenofovir disoproxil fumarate for chronic hepatitis B: a 5-year open-label follow-up study,
[cited by applicant]
Matas et al., OPTN/SRTR 2011 annual data deport: kidney,
[cited by applicant]
McAleese et al., Recruit-TandAbs: harnessing the immune system to kill cancer cells,
[cited by applicant]
Miner, The glomerular basement membrane,
[cited by applicant]
Mirmalek-Sani et al., Porcine pancreas extracellular matrix as a platform for endocrine pancreas bioengineering,
[cited by applicant]
Mitzner et al., Albumin dialysis MARS: knowledge from 10 years of clinical investigation,
[cited by applicant]
Moore et al., Application of dual affinity retargeting molecules to achieve optimal redirected T-cell killing of B-cell lymphoma,
[cited by applicant]
Moran et al., Whole-organ bioengineering: current tales of modern alchemy,
[cited by applicant]
Morrison et al., Chimeric human antibody molecules: mouse antigen-binding domains with human constant region domains,
[cited by applicant]
Nakayama et al., Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering,
[cited by applicant]
Nakayama et al., Renal tissue engineering with decellularized rhesus monkey kidneys: age-related differences,
[cited by applicant]
Nakayama et al., Tissue specificity of decellularized rhesus monkey kidney and lung scaffolds,
[cited by applicant]
Navone et al., Isolation and expansion of human and mouse brain microvascular endothelial cells,
[cited by applicant]
Neuberger et al., Recombinant antibodies possessing novel effector functions,
[cited by applicant]
Ng et al., Lineage restricted progenitors for the repopulation of decellularized heart,
[cited by applicant]
Nichols et al., Production and utilization of acellular lung scaffolds in tissue engineering,
[cited by applicant]
Orlandi et al., Cloning immunoglobulin variable domains for expression by the polymerase chain reaction,
[cited by applicant]
Orlando et al., Cell replacement strategies aiming at reconstitution of the beta cell compartment in type 1 diabetes,
[cited by applicant]
Orlando et al., Discarded human kidneys as a source of ECM scaffolds for kidney regeneration technologies,
[cited by applicant]
Orlando et al., How regenerative medicine may contribute to the achievement of an immunosuppression-free state,
[cited by applicant]
Orlando et al., Organ bioengineering and regeneration as the new Holy Grail of organ transplantation,
[cited by applicant]
Orlando et al., Pancreas transplantation for type 2 diabetes mellitus,
[cited by applicant]
Orlando et al., Production and implantation of renal extracellular matrix scaffolds from porcine kidneys as a platform for renal bioengineering investigations,
[cited by applicant]
Orlando et al., Regenerative medicine and organ transplantation: past, present and future,
[cited by applicant]
Orlando et al., Regenerative medicine as applied to general surgery,
[cited by applicant]
Orlando et al., Regenerative medicine as applied to solid organ transplantation: current status and future development,
[cited by applicant]
Orlando, Immunosuppression-free transplantation reconsidered from a regenerative medicine perspective,
[cited by applicant]
Orlando, Transplantation as a subfield of regenerative medicine. An interview by Lauren Constable,
[cited by applicant]
Ott et al., Perfusion-decellularized matrix: using nature's platform to engineer a bioartificial heart,
[cited by applicant]
Ott et al., Regeneration and orthotopic transplantation of a bioartificial lung,
[cited by applicant]
Owens et al., The genetic engineering of monoclonal antibodies,
[cited by applicant]
Pedersen et al., Comparison of surface accessible residues in human and murine immunoglobulin Fv domains. Implication for humanization of murine antibodies,
[cited by applicant]
Peloso et al., Sisyphus, the Giffen's paradox and the holy grail: time for organ transplantation to transition toward a regenerative medicine-focused type of research,
[cited by applicant]
Pereira-Sampaio et al., Cranial pole nephrectomy in the pig model: anatomic analysis of arterial injuries in tridimensional endocasts,
[cited by applicant]
Perera et al., Current status and recent advances of liver transplantation from donation after cardiac death,
[cited by applicant]
Petersen et al., Tissue-engineered lungs for in vivo implantation,
[cited by applicant]
Portner et al., T and NK cells of B cell NHL patients exert cytotoxicity against lymphoma cells following binding of bispecific tetravalent antibody CD19 x CD3 or CD19 x CD16,
[cited by applicant]
Pozzi et al., Beta1 integrin expression by podocytes is required to maintain glomerular structural integrity.
[cited by applicant]
Rahimi et al., A role for cadherin-5 in regulation of vascular endothelial growth factor receptor 2 activity in endothelial cells,
[cited by applicant]
Reiter et al., Engineering interchain disulfide bonds into conserved framework regions of Fv fragments: improved biochemical characteristics of recombinant immunotoxins containing disulfide-stabilized Fv,
[cited by applicant]
Remuzzi et al., ACE inhibition reduces glomerulosclerosis and regenerates glomerular tissue in a model of progressive renal disease,
[cited by applicant]
Remuzzi et al., Mechanisms of progression and regression of renal lesions of chronic nephropathies and diabetes,
[cited by applicant]
Reusch et al., A novel tetravalent bispecific TandAb (CD30/CD16A) efficiently recruits NK cells for the lysis of CD30+ tumor cells,
[cited by applicant]
Riechmann et al., Reshaping human antibodies for therapy,
[cited by applicant]
Roder et al., The EBV-hybridoma technique,
[cited by applicant]
Ross et al., Embryonic stem cells proliferate and differentiate when seeded into kidney scaffolds,
[cited by applicant]
Ross et al., Mouse stem cells seeded into decellularized rat kidney scaffolds endothelialize and remodel basement membranes,
[cited by applicant]
Ross et al., Regeneration and bioengineering of transplantable abdominal organs: current status and future challenges,
[cited by applicant]
Rossi et al., A new class of bispecific antibodies to redirect T cells for cancer immunotherapy,
[cited by applicant]
Salvatori et al., Extracellular matrix scaffold technology for bioartificial pancreas engineering: state of the art and future challenges,
[cited by applicant]
Salvatori et al., Semi-xenotransplantation: the regenerative medicine based-approach to immunosuppression-free transplantation and to meet the organ demand,
[cited by applicant]
Schenke-Layland et al., The use of three-dimensional nanostructures to instruct cells to produce extracellular matrix for regenerative medicine strategies,
[cited by applicant]
Schlereth et al., Eradication of tumors from a human colon cancer cell line and from ovarian cancer metastases in immunodeficient mice by a single-chain Ep-CAM-/CD3-bispecific antibody construct,
[cited by applicant]
Schlereth et al., Potent inhibition of local and disseminated tumor growth in immunocompetent mouse models by a bispecific antibody construct specific for Murine CD3,
[cited by applicant]
Shupe et al., Method for the decellularization of intact rat liver,
[cited by applicant]
Skardal et al., Tissue specific synthetic ECM hydrogels for 3-D in vitro maintenance of hepatocyte function,
[cited by applicant]
Song et al., Regeneration and experimental orthotopic transplantation of a bioengineered kidney,
[cited by applicant]
Soto-Gutierrez et al., Perspectives on whole organ assembly: moving toward transplantation on demand,
[cited by applicant]
Spenlé et al., Laminin α5 guides tissue patterning and organogenesis.
[cited by applicant]
Sullivan et al., Decellularization methods of porcine kidneys for whole organ engineering using a high-throughput system,
[cited by applicant]
Takeda et al., Construction of chimaeric processed immunoglobulin genes containing mouse variable and human constant region sequences,
[cited by applicant]
Tapias et al., Decellularized scaffolds as a platform for bioengineered organs,
[cited by applicant]
Todorovska et al., Design and application of diabodies, triabodies and tetrabodies for cancer targeting,
[cited by applicant]
Totonelli et al., Production and characterization of a rat decellularized small bowel scaffold that preserves villus-crypt architecture,
[cited by applicant]
Uygun et al., Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix,
[cited by applicant]
Verhoeyen et al., Reshaping human antibodies: grafting an antilysozyme activity,
[cited by applicant]
Vodicka et al., The miniature pig as an animal model in biomedical research,
[cited by applicant]
Wang et al., Lineage restriction of human hepatic stem cells to mature fates is made efficient by tissue-specific biomatrix scaffolds,
[cited by applicant]
Wang et al., Method for perfusion decellularization of porcine whole liver and kidney for use as a scaffold for clinical-scale bioengineering engrafts,
[cited by applicant]
Wilkin, The accelerator hypothesis: a review of the evidence for insulin resistance as the basis for type I as well as type II diabetes,
[cited by applicant]
Winter et al., Man-made antibodies,
[cited by applicant]
Winterdahl et al., Hepatic blood perfusion estimated by dynamic contrast-enhanced computed tomography in pigs: limitations of the slope method,
[cited by applicant]
Yagi et al., Whole-organ re-engineering: a regenerative medicine approach to digestive organ replacement,
[cited by applicant]
Zhang et al., Identification of proliferating human hepatic cells from human induced pluripotent stem cells,
[cited by applicant]
Zhou et al., Decellularized liver matrix as a carrier for the transplantation of human fetal and primary hepatocytes in mice,
[cited by applicant]