Vascular tissue engineering

Abstract

The invention relates to tubularized tissue, which in one embodiment is a vascular tissue made by seeding cells on the exterior surface of a scaffold and incubating of the cells so as to form a tubular tissue with the scaffold inside. In another embodiment the invention relates to a hybrid tissue. In another embodiment, the invention relates to methods of using the same for example, for enhancing blood vessel formation in a patient, for promoting angiogenesis and vasculogenesis, for replacing damaged blood vessel and for determining cellular or tissue function of an agent.

What is claimed is:  
     
         1 . A tubularized tissue made according to the steps of: 
 seeding a cell on the exterior surface of scaffold so as to obtain a scaffold encircled by cells;    incubating said scaffold encircled by said cells so as to form a tubularized tissue; thereby obtaining a tubularized tissue.    
     
     
         2 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a tubularized tissue, the tubularized tissue is a monolayer tubular tissue.  
     
     
         3 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a tubularized tissue, the tubularized tissue is a multi-layer tubular tissue.  
     
     
         4 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a tubularized tissue, the tubularized tissue is a multi-layer tubularized tissue comprising more than one type of cells.  
     
     
         5 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a tubularized tissue, said tubularized tissue is a vascularized tissue.  
     
     
         6 . The tubularized tissue of  claim 1 , whereby in the step of seeding a cell on the exterior surface of scaffold, said scaffold is in the form of a string.  
     
     
         7 . The tubularized tissue of  claim 1 , whereby in the step of seeding a cell on the exterior surface of scaffold, said scaffold is a biodegradable scaffold.  
     
     
         8 . The tubularized tissue of  claim 2 , wherein the biodegradable scaffold is a polymeric biodegradable scaffold.  
     
     
         9 . The tubularized tissue of  claim 2 , wherein the polymeric biodegradable scaffold comprising cross-linked dextran, arabinogalactan, chitosan, polyactide-glycolide, alginates, pullulan, gelatin, collagen, fibrin, fibrinogen or albumin.  
     
     
         10 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a scaffold, the diameter of said scaffold is 4-500 um.  
     
     
         11 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a scaffold, said scaffold is degradable upon exposure to predetermined environmental conditions.  
     
     
         12 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a scaffold, said scaffold is a non-biodegradable scaffold.  
     
     
         13 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a scaffold, said scaffold comprises macromolecules necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         14 . The tubularized tissue of  claim 13 , wherein the macromolecules necessary for cell growth is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         15 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a scaffold, said scaffold contains also a therapeutic agent.  
     
     
         16 . The tubularized tissue of  claim 15 , wherein said therapeutic agent is an anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         17 . The tubularized tissue of  claim 15 , wherein said therapeutic agent is a nucleic acid sequence which encodes an anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         18 . The tubularized tissue of  claim 1 , wherein said cell is engineered to express a therapeutic agent and/or a macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         19 . The tubularized tissue of  claim 18 , wherein said therapeutic agent is anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         20 . The tubularized tissue of  claim 18 , wherein said macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         21 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a cell, said cell is a mesenchymal stem cell, mesodermal progenitor cell, endothelial precursor cell or neonatal dermal micro vascular endothelial cells.  
     
     
         22 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a cell, said cell is capable of differentiating into an endotelial cell.  
     
     
         23 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a cell, said cell is isolated from peripheral blood.  
     
     
         24 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a cell, said cell is isolated from mammalian artery, vein or capillary.  
     
     
         25 . The tubularized tissue of  claim 1 , whereby in the step of obtaining a cell said cell is genetically engineered to express a reporter gene.  
     
     
         26 . The tubularized tissue of  claim 25 , wherein the reporter gene is a fluorescent protein, Luciferase or b-gal.  
     
     
         27 . The tubularized tissue of  claim 1 , wherein said step of growing scaffold encircled by cells so as to form a tubularized tissue is by the use of a bioreactor.  
     
     
         28 . The tubularized tissue of  claim 1 , wherein the tubularized tissue is a single vessel, composition of several vessels or a capillary network.  
     
     
         29 . A method of maturing cells into a tubularized tissue comprising the following steps: 
 seeding a cell on the exterior surface of scaffold so as to obtain a scaffold encircled by cells;    incubating said scaffold encircled by said cells so as to form a tubularized tissue; thereby maturing cells into a tubularized tissue.    
     
     
         30 . The method of  claim 29 , wherein said tubularized tissue is a monolayer tubular tissue.  
     
     
         31 . The method of  claim 29 , wherein said tubularized tissue is a multilayer tubular tissue.  
     
     
         32 . The method of  claim 29 , wherein said tubularized tissue is a multi-layer tubularized tissue comprising more than one type of cells.  
     
     
         33 . The method of  claim 29 , wherein said tubularized tissue is a vascular tissue.  
     
     
         34 . The method of  claim 29 , wherein said scaffold is in the form of a string.  
     
     
         35 . The method of  claim 29 , wherein said scaffold is a biodegradable scaffold.  
     
     
         36 . The method of  claim 35 , wherein said biodegradable scaffold is a polymeric biodegradable scaffold.  
     
     
         37 . The method of  claim 36 , wherein said polymeric biodegradable scaffold comprising cross-linked dextran, arabinogalactan, chitosan, polyactide-glycolide, alginates, pullulan, gelatin, collagen, fibrin, fibrinogen or albumin.  
     
     
         38 . The method of  claim 29 , wherein the diameter of said scaffold is 4-500 um.  
     
     
         39 . The method of  claim 29 , wherein said scaffold is degradable upon exposure to predetermined environmental conditions.  
     
     
         40 . The method of  claim 29 , wherein said scaffold is a non-biodegradable scaffold.  
     
     
         41 . The method of  claim 29 , wherein said scaffold comprises macromolecules necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         42 . The method of  claim 41 , wherein said macromolecules necessary for cell growth is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         43 . The method of  claim 29 , wherein said scaffold contains also a therapeutic agent.  
     
     
         44 . The method of  claim 43 , wherein said therapeutic agent is an anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         45 . The method of  claim 29 , wherein said cell is engineered to express a therapeutic agent and/or a macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         46 . The method of  claim 45 , wherein said therapeutic agent is anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         47 . The method of  claim 45 , wherein said macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         48 . The method of  claim 29 , wherein said cell is a mesenchymal stem cell, mesodermal progenitor cell, endothelial precursor cell or neonatal dermal micro vascular endothelial cells.  
     
     
         49 . The method of  claim 29 , wherein said cell is capable of differentiating into an endotelial cell.  
     
     
         50 . The method of  claim 29 , wherein said cell is isolated from peripheral blood.  
     
     
         51 . The method of  claim 29 , wherein said cell is isolated from mammalian artery, vein or capillary.  
     
     
         52 . The method of  claim 29 , wherein said cell is genetically engineered to express a reporter gene.  
     
     
         53 . The method of  claim 52 , wherein the reporter gene is a fluorescent protein, Luciferase or b-gal.  
     
     
         54 . The method of  claim 29 , wherein said step of growing scaffold encircled by cells so as to form a tubularized tissue is by the use of a bioreactor.  
     
     
         55 . The method of  claim 29 , wherein tubularized tissue is a single vessel, composition of several vessels or a capillary network.  
     
     
         56 . A method of enhancing blood vessel formation in a patient in need thereof, comprising the following steps: 
 selecting the patient in need thereof;    isolating a cell from the patient;    seeding a cell on the exterior surface of scaffold so as to obtain a scaffold encircled by cells;    incubating said scaffold encircled by said cells so as to form a tubularized tissue; and readministering said tubularized tissue to the patient in need thereof, thereby enhancing blood vessel formation in a patient in need thereof.    
     
     
         57 . The method of  claim 56 , wherein said tubularized tissue is a monolayer tubular tissue.  
     
     
         58 . The method of  claim 56 , wherein said tubularized tissue is a multi-layer tubular tissue.  
     
     
         59 . The method of  claim 56 , wherein said tubularized tissue is a multi-layer tubularized tissue comprising more than one type of cells.  
     
     
         60 . The method of  claim 56 , wherein said tubularized tissue is a vascular tissue.  
     
     
         61 . The method of  claim 56 , wherein said scaffold is in the form of a string.  
     
     
         62 . The method of  claim 56 , wherein said scaffold is a biodegradable scaffold.  
     
     
         63 . The method of  claim 62 , wherein said biodegradable scaffold is a polymeric biodegradable scaffold.  
     
     
         64 . The method of  claim 63 , wherein said polymeric biodegradable scaffold comprising cross-linked dextran, arabinogalactan, chitosan, polyactide-glycolide, alginates, pullulan, gelatine, collagen, fibrin, fibrinogen or albumin.  
     
     
         65 . The method of  claim 56 , wherein the diameter of said scaffold is 4-10000 um.  
     
     
         66 . The method of  claim 56 , wherein said scaffold is degradable upon exposure to predetermined environmental conditions.  
     
     
         67 . The method of  claim 56 , wherein said scaffold is a non-biodegradable scaffold.  
     
     
         68 . The method of  claim 56 , wherein said scaffold comprises macromolecules necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         69 . The method of  claim 68 , wherein said macromolecules necessary for cell growth is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         70 . The method of  claim 56 , wherein said scaffold contains also a therapeutic agent.  
     
     
         71 . The method of  claim 70 , wherein said therapeutic agent is an anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         72 . The method of  claim 56 , wherein said cell is engineered to express a therapeutic agent and/or a macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         73 . The method of  claim 72 , wherein said therapeutic agent is anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         74 . The method of  claim 72 , wherein said macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         75 . The method of  claim 56 , wherein said cell is a mesenchymal stem cell, mesodermal progenitor cell, endothelial precursor cell or neonatal dermal micro vascular endothelial cells (foreskin).  
     
     
         76 . The method of  claim 56 , wherein said cell is capable of differentiating into an endotelial cell.  
     
     
         77 . The method of  claim 56 , wherein said cell is isolated from peripheral blood.  
     
     
         78 . The method of  claim 56 , wherein said cell is isolated from mammalian artery, vein or capillary.  
     
     
         79 . The method of  claim 56 , wherein said cell is genetically engineered to express a reporter gene.  
     
     
         80 . The method of  claim 79 , wherein the reporter gene is a fluorescent protein, Luciferase or b-gal.  
     
     
         81 . The method of  claim 56 , wherein said step of growing scaffold encircled by cells so as to form a tubularized tissue is by the use of a bioreactor.  
     
     
         82 . The method of  claim 56 , wherein the tubularized tissue is a single vessel, composition of several vessels or a capillary network.  
     
     
         83 . A method of regulating angiogenesis comprising administering to said patient host an effective amount of the tubularized tissue of  claim 1 , so as to promoting angiogenesis.  
     
     
         84 . A method of regulating vasculogenesis in a tissue comprising administering to said patient host an effective amount of the tubularized tissue of  claim 1 , so as to promoting vasculogenesis.  
     
     
         85 . A method of treating hypoxia, avascular necrosis, foot ulcer or gangrene result from diabetes, athersclerosis, stroke, wound, fracture of bone, comprising administering to said patient host an effective amount of the tubularized tissue of  claim 1  so as to increasing the vascularity of a tissue thereby treating.  
     
     
         86 . A method of replacing a damaged blood vessel of a patient comprising the step of administering to the patient an effective amount of the tubularized tissue, thereby replacing damaged blood vessels.  
     
     
         87 . The method of  claim 86 , wherein the blood vessel is a thick blood vessels or a thin blood vessel.  
     
     
         88 . A method for inducing the formation of new blood vessels in an ischemic tissue in a patient in need thereof, comprising: 
 administering to said patient host an effective amount of the tubularized tissue of  claim 1 , thereby inducing the formation of new blood vessels.    
     
     
         89 . The method of  claim 88 , wherein said patient is in need of treatment for cerebrovascular ischemia, renal ischemia, pulmonary ischemia, limb ischemia, ischemic cardiomyopathy and myocardial ischemia.  
     
     
         90 . A method of enabling ex vivo organ engineering by introducing the tubularized tissue of  claim 1  to an engineered tissue so as to enable a constant oxygen supply to said engineered tissue.  
     
     
         91 . A method of engrafting in a patient in need thereof, comprising administering to said patient host an effective amount of the tubularized tissue of  claim 1 , thereby engrafting in a patient in need thereof.  
     
     
         92 . A method of determining cellular or tissue function of an agent comprising the following steps: 
 obtaining the tubularized tissue of  claim 1;     contacting said tubularized tissue with an agent, so as to obtain a tubularized tissue contacted with an agent;    comparing cellular or tissue function of said tubularized tissue contacted with an agent, to a tubularized tissue, thereby determining cellular or tissue function of the agent.    
     
     
         93 . The method of  claim 92 , wherein the cellular or tissue function is proliferation, apoptosis, toxicity and differentiation.  
     
     
         94 . A method of screening candidate genes which are involved in cellular or tissue function, said method comprising the steps of: 
 obtaining the tubularized tissue of  claim 1;     obtaining mRNA from said tubularized tissue and said cell;    synthesizing cDNA from said mRNA; amplifying said cDNA-hybrid, so as to obtain an amplified product;    detecting said amplified product; and comparing said amplified products from said tubularized tissue and said cell, thereby identifying candidate nucleic acid sequence which is involved in the cellular or tissue function.    
     
     
         95 . An ex-vivo system for determining cellular or tissue function of an agent according to the method of  claim 92 .  
     
     
         96 . An ex-vivo system for screening candidate genes, which are involved in cellular or tissue according to the method of  claim 94 .  
     
     
         97 . A hybrid tissue made according to the steps of: 
 seeding a cell that will form a first tissue type on the exterior surface of a branched scaffold so as to obtain a scaffold encircled by cells with spaces in between;    adding into said spaces in between, a cell that will form a second tissue type; and    incubating so as to obtain a hybrid tissue.    
     
     
         98 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a hybrid tissue, the hybrid tissue is a monolayer tubular tissue.  
     
     
         99 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a hybrid tissue, the hybrid tissue is a multilayer tubular tissue.  
     
     
         100 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a hybrid tissue, the hybrid tissue is a multilayer hybrid tissue comprising more than one type of cells.  
     
     
         101 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a hybrid tissue, said hybrid tissue is a vascularized tissue.  
     
     
         102 . The hybrid tissue of  claim 97 , whereby in the step of seeding a cell on the exterior surface of scaffold, said scaffold is in the form of a string.  
     
     
         103 . The hybrid tissue of  claim 97 , whereby in the step of seeding a cell on the exterior surface of scaffold, said scaffold is a biodegradable scaffold.  
     
     
         104 . The hybrid tissue of  claim 103 , wherein the biodegradable scaffold is a polymeric biodegradable scaffold.  
     
     
         105 . The hybrid tissue of  claim 104 , wherein the polymeric biodegradable scaffold comprising cross-linked dextran, arabinogalactan, chitosan, polyactide-glycolide, alginates, pullulan, gelatine, collagen, fibrin, fibrinogen or albumin.  
     
     
         106 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a scaffold, the diameter of said scaffold is 4-500 um.  
     
     
         107 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a scaffold, said scaffold is degradable upon exposure to predetermined environmental conditions.  
     
     
         108 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a scaffold, said scaffold is a non-biodegradable scaffold.  
     
     
         109 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a scaffold, said scaffold comprises macromolecules necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         110 . The hybrid tissue of  claim 109 , wherein the macromolecules necessary for cell growth is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         111 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a scaffold, said scaffold contains also a therapeutic agent.  
     
     
         112 . The hybrid tissue of  claim 111 , wherein said therapeutic agent is an anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         113 . The hybrid tissue of  claim 111 , wherein said therapeutic agent is an a nucleic acid sequence which encodes an anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         114 . The hybrid tissue of  claim 97 , wherein said cell is engineered to express a therapeutic agent and/or a macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof.  
     
     
         115 . The hybrid tissue of  claim 114 , wherein said therapeutic agent is anti-infective, a hormone, an analgesic, an anti-inflammatory agent, a chemotherapeutic agent, an anti-rejection agent, a prostaglandin, RGD peptide and combinations thereof.  
     
     
         116 . The hybrid tissue of  claim 114 , wherein said macromolecule necessary for cell growth, morphogenesis, differentiation, or tissue building and combinations thereof is a bone morphogenic protein, a bone morphogenic-like protein, an epidermal growth factor, a fibroblast growth factor, a platelet derived growth factor, an insulin like growth factor, a transforming growth factor, a vascular endothelial growth factor, Ang1, PIGF and combinations thereof.  
     
     
         117 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a cell, said cell is a mesenchymal stem cell, mesodermal progenitor cell, endothelial precursor cell or neonatal dermal micro vascular endothelial cells.  
     
     
         118 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a cell, said cell is capable of differentiating into an endotelial cell.  
     
     
         119 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a cell, said cell is isolated from peripheral blood.  
     
     
         120 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a cell, said cell is isolated from mammalian artery, vein or capillary.  
     
     
         121 . The hybrid tissue of  claim 97 , whereby in the step of obtaining a cell said cell is genetically engineered to express a reporter gene.  
     
     
         122 . The hybrid tissue of  claim 121 , wherein the reporter gene is a fluorescent protein, Luciferase or b-gal.  
     
     
         123 . The hybrid tissue of  claim 97 , wherein said step of growing scaffold encircled by cells so as to form a hybrid tissue is by the use of a bioreactor.  
     
     
         124 . The hybrid tissue of  claim 97 , wherein the hybrid tissue is a single vessel, composition of several vessels or a capillary network.