Vascular tissue engineering

• US 20030068817A1
• Filed: 09/05/2002
• Published: 04/10/2003
• Est. Priority Date: 04/30/2001
• Status: Abandoned Application

• Current Assignee: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. (Hebrew University of Jerusalem)
• Original Assignee: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. (Hebrew University of Jerusalem)
• Inventors: Gafni, Yossi, Pelled, Gadi, Gazit, Dan, Turgeman, Gadi
• Application Number:
  US10/234,707
• Publication Number:
  US 20030068817A1
• Time in Patent Office: Days
• US Class Current: 435/325
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2300/426 Immunomodulating agents, i.e. cytokines, interleukins, interferons
• CPC Class Codes: A61L 2300/604 Biodegradation
• CPC Class Codes: A61L 2430/36 for embolization or occlusion, e.g. vaso-occlusive compositions or devices
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/22 Polypeptides or derivatives thereof , e.g. degradation products
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3625 Vascular tissue, e.g. heart valves
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3886 comprising two or more cell types
• CPC Class Codes: A61L 27/3891 as distinct cell layers
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/40 Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
• CPC Class Codes: A61L 27/48 with macromolecular fillers
• CPC Class Codes: A61L 27/507 for artificial blood vessels apparatus for applying cells on a blood vessel prosthesis A61F2/062
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: C08L 67/04 Polyesters derived from hydroxycarboxylic acids, e.g. lactones C08L67/06 takes precedence
• CPC Class Codes: C08L 71/02 : Polyalkylene oxides
• CPC Class Codes: C12M 25/14 : Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12N 2501/155 : Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2502/28 : Vascular endothelial cells
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 2533/40 : Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/54 : Collagen; Gelatin
• CPC Class Codes: C12N 2533/70 : Polysaccharides
• CPC Class Codes: C12N 2533/72 : Chitin, chitosan
• CPC Class Codes: C12N 5/0068 : General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0662 : Stem cells

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.

Claims

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.

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.

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.

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.

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.

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.