DECELLULARIZATION AND RECELLULARIZATION OF ORGANS AND TISSUES
The invention provides for methods and materials to decellularize a solid organ and to recellularize such a decellularized organ to thereby generate a solid organ.
1 . A composition that comprises:
an at least partially decellularized extracellular matrix of a pig, a bovine, a sheep, or a canine organ or portion thereof, wherein said at least partially decellularized extracellular matrix is cannulated and: i) comprises at least a portion of an intact exterior surface, ii) upon perfusion, retains a majority of fluid perfused into a vascular tree of said at least partially decellularized extracellular matrix, or iii) a combination of i) and ii).
2 . The composition of claim 1 , wherein said organ or said portion thereof is from said pig.
3 . The composition of claim 1 , wherein said organ or said portion thereof is a heart, a liver, a lung, a skeletal muscle, a bowel, a stomach, a bone, a brain, a pancreas, a spleen, a kidney, a uterus, a bladder, or a portion of any of these.
4 . The composition of claim 3 , wherein said organ or said portion thereof is said lung or said portion of said lung, wherein said portion of said lung is a trachea.
5 . The composition of claim 3 , wherein said organ or portion thereof is said kidney or said portion thereof.
6 . The composition of claim 3 , wherein said organ or said portion thereof is said liver or said portion thereof.
7 . The composition of claim 3 , wherein said organ or said portion thereof is said heart or said portion thereof, wherein said portion of said heart is selected from the group consisting of: an aortic valve, a mitral valve, a pulmonary valve, a tricuspid valve, a right atrium, a left atrium, a right ventricle, a left ventricle, a cardiac patch, septum, a coronary vessel, a pulmonary artery, and a pulmonary vein.
8 . The composition of claim 1 , wherein said at least partially decellularized matrix further comprises an exogenous population of cells that comprises at least one cell that is xenogenic to said at least partially decellularized extracellular matrix.
9 . The composition of claim 8 , wherein said cell that is xenogeneic is human.
10 . A bioreactor, that comprises:
(a) an at least partially decellularized extracellular matrix of a pig, a bovine, a sheep, or a canine organ or portion thereof, wherein said at least partially decellularized extracellular matrix: i) comprises at least a portion of an intact exterior surface, ii) upon perfusion, retains a majority of fluid perfused into a vascular tree of said at least partially decellularized extracellular matrix, or iii) a combination of i) and ii); and
(b) a component selected from the group consisting of: a cannula, a perfusion apparatus, a holding container, tubing, a pump, a sensor, a thermometer, an electrode, a valve, a balloon, a pacemaker, a thermostat, and any combination thereof.
11 . The bioreactor of claim 10 , wherein said bioreactor comprises at least two of said components.
12 . The bioreactor of claim 10 , wherein said at least partially decellularized organ or portion thereof is a heart, a kidney, a liver, a pancreas, a spleen, a bowel, a skeletal muscle, a bone, a brain, a uterus, a bladder, or a lung.
13 . The bioreactor of claim 12 , wherein said at least partially decellularized extracellular matrix of said organ or portion thereof is said heart, or a portion of said heart selected from the group consisting of: an aortic valve, a mitral valve, a pulmonary valve, a tricuspid valve, a right atrium, a left atrium, a right ventricle, a left ventricle, a cardiac patch, septum, a coronary vessel, a pulmonary artery, a pulmonary vein, and any combination thereof.
14 . The bioreactor of claim 11 , wherein said component is said cannula, and wherein a vessel, a duct, a cavity, or a combination thereof of said at least partially decellularized extracellular matrix of said organ or portion thereof is cannulated.
15 . The bioreactor of claim 10 , wherein said at least partially decellularized extracellular matrix of said organ or portion thereof further comprises an exogenous population of cells.
16 . The bioreactor of claim 15 , wherein said exogenous population of cells comprises a xenogeneic cell to said at least partially decellularized extracellular matrix.
17 . The bioreactor of claim 16 , wherein said xenogeneic cell is human.
18 . The bioreactor of claim 17 , wherein said human cell is: an embryonic stem cells or a subset thereof, an umbilical cord cell or a subset thereof, a bone marrow cell or a subset thereof, a peripheral blood cell or a subset thereof, an adult-derived stem cell or a subset thereof, a progenitor cell or a subset thereof, a tissue-derived stem or progenitor cell or a subset thereof, a mesenchymal stem cell (MSC) or a subset thereof, a skeletal muscle-derived stem or progenitor cell or a subset thereof, a multipotent adult progenitor cell (MAPC) or a subset thereof, a cardiac stem cell (CSC) or a subset thereof, or a multipotent adult cardiac-derived stem cell or a subset thereof.
19 . The bioreactor of claim 15 , wherein said exogenous population of cells comprises a heart cell, and wherein said heart cell is a cardiac fibroblast, a cardiac microvasculature endothelial cell, or an aortic endothelial cell.
20 . The bioreactor of claim 10 , wherein said at least partially decellularized extracellular matrix further comprises a solution, and wherein said solution comprises an enzyme, a detergent, a compound, an immune modulating agent, a coagulation cascade modifier, a hormone, a supplement, an antibiotic, an anti-fungal, or an anti-microbial.
21 . The bioreactor of claim 16 , wherein said xenogeneic cell is human leukocyte antigen (HLA)-matched to a subject in need thereof.
22 . The bioreactor of claim 16 , wherein said xenogeneic cell is non-human leukocyte antigen (HLA) matched to a subject in need thereof.
23 . A method, comprising
introducing a population of cells into an at least partially decellularized extracellular matrix of a pig, a bovine, a sheep, or a canine organ or portion thereof, wherein said at least partially decellularized extracellular matrix: i) comprises at least a portion of an intact exterior surface, ii) upon perfusion, retains a majority of fluid perfused into a vascular tree of said at least partially decellularized extracellular matrix, or iii) a combination of i) and ii), and wherein at least a portion of said population of cells distribute in at least a portion of said at least partially decellularized extracellular matrix.
24 . The method of claim 23 , wherein said introducing said population of cells comprises injected perfusing or injecting said population of cells into said at least partially decellularized extracellular matrix.
25 . The method of claim 23 , wherein said at least partially decellularized extracellular matrix or portion thereof is a heart, a kidney, a liver, a spleen, a pancreas, a skeletal muscle, a bowel, a stomach, a bone, a brain, a uterus, a bladder, a lung, or a portion of any of these.
26 . A method, comprising:
perfusing a cannulated organ or portion thereof with a medium via said cannula, so as to generate an at least partially decellularized extracellular matrix, wherein said at least partially decellularized extracellular matrix: i) comprises at least a portion of an intact exterior surface, ii) upon perfusion, retains a majority of fluid perfused into a vascular tree of said at least partially decellularized extracellular matrix, or iii) a combination of i) and ii).
27 . The method of claim 26 , wherein said medium is a first cellular disruption medium, and wherein said first cellular disruption medium comprises a detergent.
28 . The method of claim 26 , wherein said cannulated organ or portion thereof is a heart, a kidney, a liver, a spleen, a pancreas, a skeletal muscle, a bowel, a stomach, a bone, a brain, a uterus, a bladder, or a lung.
29 . The method of claim 26 , wherein said perfusing is antegrade.
30 . The method of claim 26 , wherein said perfusing is retrograde.
31 . The method of claim 27 , wherein said detergent is selected from the group consisting of Sodium Dodecyl Sulfate (SDS), Polyethylene Glycol (PEG), and Triton X.
32 . The method of claim 27 , further comprising perfusing said cannulated organ or portion thereof with a second cellular disruption medium via said cannula.
33 . The method of claim 32 , wherein said detergent in said first cellular disruption medium is an anionic detergent and wherein said second cellular disruption medium comprises an ionic detergent.
34 . The method of claim 33 , wherein said anionic detergent is SDS and wherein said ionic detergent is Triton X.
35 . (canceled)
36 . An at least partially decellularized vascular mammalian tissue or portion thereof, that comprises an at least partially decellularized extracellular matrix of an organ or portion thereof, wherein said at least partially decellularized extracellular matrix is cannulated and: i) comprises at least a portion of an intact exterior surface, ii) upon perfusion, retains a majority of fluid perfused into a vascular tree of said at least partially decellularized extracellular matrix, or iii) a combination of i) and ii)
37 . The partially decellularized tissue of claim 36 , wherein said tissue or portion thereof is cadaveric.
38 . The partially decellularized tissue of claim 36 , wherein said tissue or portion thereof is a spinal cord, a trachea, or a joint, or portion thereof.
39 . A bioreactor, that comprises:
an at least partially decellularized extracellular matrix of an avascular tissue or portion thereof, wherein said at least partially decellularized extracellular matrix comprises at least a portion of an intact exterior surface; and
(b) a component selected from the group consisting of: a cannula, a perfusion apparatus, a holding container, tubing, a pump, a sensor, a thermometer, an electrode, a valve, a balloon, a pacemaker, a thermostat, and any combination thereof exterior surface.
40 . The bioreactor of claim 39 , wherein said avascular tissue or portion thereof is cartilage, a cornea, or a portion thereof.
41 . bioreactor of claim 39 , wherein said avascular tissue or portion thereof is cadaveric.
42 . The composition of claim 1 , wherein said at least partially decellularized extracellular matrix, upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix and wherein said vascular tree is substantially intact.
43 . The bioreactor of claim 10 , wherein said at least partially decellularized extracellular matrix, upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix, and wherein said vascular tree is substantially intact.
44 . The method of claim 23 , wherein said at least partially decellularized extracellular matrix, upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix, and wherein said vascular tree is substantially intact.
45 . The method of claim 26 , wherein said at least partially decellularized extracellular matrix, upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix, and wherein said vascular tree is substantially intact.
46 . The partially decellularized tissue of claim 36 , wherein said at least partially decellularized extracellular matrix, upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix, and wherein said vascular tree is substantially intact.
47 . The composition of claim 1 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface.
48 . The bioreactor of claim 10 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface.
49 . The method of claim 23 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface.
50 . The method of claim 26 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface.
51 . The partially decellularized tissue of claim 36 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface.
52 . The composition of claim 1 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface, and upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix and wherein said vascular tree is substantially intact.
53 . The bioreactor of claim 10 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface, and upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix and wherein said vascular tree is substantially intact.
54 . The method of claim 23 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface, and upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix and wherein said vascular tree is substantially intact.
55 . The method of claim 26 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface, and upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix and wherein said vascular tree is substantially intact.
56 . The partially decellularized tissue of claim 36 , wherein said at least partially decellularized extracellular matrix comprises at least a portion of the intact exterior surface, and upon perfusion, retains said majority of said fluid perfused into said vascular tree of said at least partially decellularized extracellular matrix and wherein said vascular tree is substantially intact.