Cartilage and bone repair and regeneration using postpartum-derived cells

• US 20050019865A1
• Filed: 06/25/2004
• Published: 01/27/2005
• Est. Priority Date: 06/27/2003
• Status: Active Grant

• Current Assignee: Depuy Synthes Products Incorporated (Johnson & Johnson)
• Original Assignee: Depuy Synthes Products Incorporated (Johnson & Johnson)
• Inventors: Harmon, Alexander M., Messina, Darin J., Mistry, Sanjay, Gosiewska, Anna, Seyda, Agnieszka, Kihm, Anthony J., Dhanaraj, Sridevi, Harris, Ian Ross, Yi, Chin-Feng, Wang, Ziwei
• Granted Patent: US 11,191,789 B2
• Time in Patent Office: Days
• US Class Current: 435/69.100
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: A61K 38/18 Growth factors; Growth regulators
• CPC Class Codes: A61K 38/1808 Epidermal growth factor [EGF] urogastrone
• CPC Class Codes: A61K 38/1825 Fibroblast growth factor [FGF]
• CPC Class Codes: A61K 38/1833 Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
• CPC Class Codes: A61K 38/1841 Transforming growth factor [TGF]
• CPC Class Codes: A61K 38/185 Nerve growth factor [NGF]; Brain derived neurotrophic factor [BDNF]; Ciliary neurotrophic factor [CNTF]; Glial derived neurotrophic factor [GDNF]; Neurotrophins, e.g. NT-3
• CPC Class Codes: A61K 38/1858 Platelet-derived growth factor [PDGF]
• CPC Class Codes: A61K 38/1866 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: A61K 38/1891 Angiogenesic factors; Angiogenin
• CPC Class Codes: A61K 38/19 Cytokines; Lymphokines; Interferons
• CPC Class Codes: A61K 38/204 IL-6
• CPC Class Codes: A61K 38/2053 IL-8
• CPC Class Codes: A61K 38/27 Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 13/12 : of the kidneys
• CPC Class Codes: A61P 17/02 : for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 : Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 : for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/08 : for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 19/10 : for osteoporosis
• CPC Class Codes: A61P 21/00 : Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 : Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 : for peripheral neuropathies
• CPC Class Codes: A61P 25/14 : for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 : Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 : for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 27/06 : Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/06 : Free radical scavengers or antioxidants
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2500/32 : Amino acids
• CPC Class Codes: C12N 2500/34 : Sugars
• CPC Class Codes: C12N 2500/44 : Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2500/90 : Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2500/95 : Protein-free medium and culture conditions
• CPC Class Codes: C12N 2501/12 : Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/21 : Chemokines, e.g. MIP-1, MIP-2, RANTES, MCP, PF-4
• CPC Class Codes: C12N 2501/23 : Interleukins [IL]
• CPC Class Codes: C12N 2502/02 : embryonic cells
• CPC Class Codes: C12N 2506/02 : from embryonic cells
• CPC Class Codes: C12N 2506/03 : from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2509/00 : Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2533/50 : Proteins
• CPC Class Codes: C12N 5/0605 : Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0606 : Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0607 : Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696

Abstract

Cells derived from postpartum tissue and methods for their isolation and induction to differentiate to cells of a chondrogenic or osteogenic phenotype are provided by the invention. The invention further provides cultures and compositions of the postpartum-derived cells and products related thereto. The postpartum-derived cells of the invention and products related thereto have a plethora of uses, including but not limited to research, diagnostic, and therapeutic applications, for example, in the treatment of bone and cartilage conditions.

Claims

1. A postpartum-derived cell comprising a cell derived from human postpartum tissue substantially free of blood, wherein said cell is capable of self-renewal and expansion in culture and has the potential to differentiate into a cell of an osteogenic or chondrogenic phenotype;

wherein said cell requires L-valine for growth;

wherein said cell is capable of growth in about 5% to about 20% oxygen;

wherein said cell further comprises at least one of the following characteristics;

(a) production of at least one of granulocyte chemotactic protein 2 (GCP-2), reticulon 1, tissue factor, vimentin, and alpha-smooth muscle actin;

(b) lack of production of at least one of GRO-alpha or oxidized low density lipoprotein receptor, as detected by flow cytometry;

(c) production of at least one of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A,B,C;

(d) lack of production of at least one of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, and HLA-DR,DP,DQ, as detected by flow cytometry;

(e) expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is increased for at least one of interleukin 8;

reticulon 1;

chemokine (C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha);

chemokine (C-X-C motif) ligand 6 (granulocyte chemotactic protein

     2);

chemokine (C-X-C motif) ligand 3; and

tumor necrosis factor, alpha-induced protein 3 or expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is increased for at least one of C-type lectin superfamily member A2, Wilms tumor 1, aldehyde dehydrogenase 1 family member A2, renin, oxidized low density lipoprotein receptor 1, protein kinase C zeta, clone IMAGE;

4179671, hypothetical protein DKFZp564F013, downregulated in ovarian cancer 1, and clone DKFZp547K1113;

(f) expression, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, is reduced for at least one of;

short stature homeobox 2;

heat shock 27 kDa protein 2;

chemokine (C-X-C motif) ligand 12 (stromal cell-derived factor

     1);

elastin;

cDNA DKFZp586M2022 (from clone DKFZp586M2022);

mesenchyme homeobox 2;

sine oculis homeobox homolog 1;

crystallin, alpha B;

dishevelled associated activator of morphogenesis 2;

DKFZP586B2420 protein;

similar to neuralin 1;

tetranectin;

src homology three (SH3) and cysteine rich domain;

B-cell translocation gene 1, anti-proliferative;

cholesterol 25-hydroxylase;

runt-related transcription factor 3;

hypothetical protein FLJ23191;

interleukin 11 receptor, alpha;

procollagen C-endopeptidase enhancer;

frizzled homolog 7;

hypothetical gene BC008967;

collagen, type VIII, alpha 1;

tenascin C;

iroquois homeobox protein 5;

hephaestin;

integrin, beta 8;

synaptic vesicle glycoprotein 2;

cDNA FLJ12280 fis, clone MAMMA1001744;

cytokine receptor-like factor 1;

potassium intermediate/small conductance calcium-activated channel, subfamily N, member 4;

integrin, alpha 7;

DKFZP586L151 protein;

transcriptional co-activator with PDZ-binding motif (TAZ);

sine oculis homeobox homolog 2;

KIAA1034 protein;

early growth response 3;

distal-less homeobox 5;

hypothetical protein FLJ20373;

aldo-keto reductase family 1, member C3 (3-alpha hydroxysteroid dehydrogenase, type II);

biglycan;

fibronectin 1;

proenkephalin;

integrin, beta-like 1 (with EGF-like repeat domains);

cDNA clone EUROIMAGE 1968422;

EphA3;

KIAA0367 protein;

natriuretic peptide receptor C/guanylate cyclase C (atrionatriuretic peptide receptor C);

hypothetical protein FLJ14054;

cDNA DKFZp564B222 (from clone DKFZp564B222);

vesicle-associated membrane protein 5;

EGF-containing fibulin-like extracellular matrix protein 1;

BCL2/adenovirus E1B 19 kDa interacting protein 3-like;

AE binding protein 1;

cytochrome c oxidase subunit VIIa polypeptide 1 (muscle);

neuroblastoma, suppression of tumorigenicity 1; and

insulin-like growth factor binding protein 2, 36 kDa;

(g) secretion of at least one of monocyte chemotactic protein-1, interleukin (IL)-6, IL-8, granulocyte chemotactic protein-2, hepatocyte growth factor, keratinocyte growth factor, fibroblast growth factor, heparin binding-epidermal growth factor, brain derived neurotrophic factor, thrombopoietin, macrophage inflammatory protein (MIP)-1a, RANTES, and tissue inhibitor of matrix metalloprotease 1;

(h) lack of secretion of at least one of transforming growth factor-beta2, angiopoetin-2, platelet derived growth factor-bb, MIP1b, 1309, macrophage-derived chemokine, and vascular endothelial growth factor, as detected by ELISA; and

(i) the ability to undergo at least 40 population doublings in culture.

2. The cell of claim 1 which has been isolated from a post-partum placenta or fragment thereof by enzymatic dissociation with at least one of a matrix metalloprotease, a neutral protease, and a mucolytic enzyme that digests hyaluronic acid.

3. A method of inducing differentiation of a postpartum-derived cell of claim 1 to a chondrogenic phenotype comprising exposing said cell to one or more chondrogenic differentiation-inducing agents.

4. The method of claim 3 wherein said chondrogenic differentiation-inducing agent comprises at least one of transforming growth factor-beta3 (TGFbeta3) and growth and differentiation factor-5 (GDF-5).

5. The method of claim 3 further comprising culturing the cell in chondrogenic medium.

6. The method of claim 5 wherein said chondrogenic medium comprises Dulbecco'"'"'s modified Eagle'"'"'s medium, L-glutamine, sodium pyruvate, L-proline, dexamethasone, L-ascorbic acid, insulin, transferrin, selenium, and an antibiotic agent.

7. The method of claim 6 wherein said chondrogenic medium further comprises at least one of collagen and sodium hydroxide.

8. The method of claim 3 further comprising evaluating differentiation of said cell by a pellet culture assay.

9. The method of claim 3 further comprising evaluating differentiation of said cell by detecting the presence of a glycosaminoglycan or collagen.

10. The method of claim 9 wherein said step of evaluating comprises staining said cell with Safranin-O or hematoxylin/eosin.

11. The cell produced by the method of claim 3.

12. A method of inducing differentiation of a postpartum-derived cell of claim 1 to an osteogenic phenotype comprising exposing said cell to one or more osteogenic differentiation-inducing agents.

13. The method of claim 12 wherein said differentiation-inducing agent comprises at least one of bone morphogenic protein (BMP)-2, BMP-4, and transforming growth factor-beta1.

14. The method of claim 12 further comprising culturing said cell in osteogenic medium.

15. The method of claim 14 wherein said osteogenic medium comprises Dulbecco'"'"'s modified Eagle'"'"'s medium-low glucose, serum, beta-glycerophosphate, dexamethasone, ascorbic phosphate salt, and at least one antibiotic or antimycotic agent.

16. The method of claim 12 further comprising evaluating said differentiation by detecting an osteogenic lineage-specific marker.

17. The method of claim 16 wherein said marker is osteocalcin, bone sialoprotein, or alkaline phosphatase.

18. The method of claim 12 further comprising detecting said differentiation by measuring mineralization.

19. The method of claim 18 wherein said step of detecting comprises von Kossa staining.

20. The cell produced according to the method of claim 12.

21. A cell population comprising the postpartum-derived cell of claim 1.

22. The cell population of claim 21 wherein said cell population is substantially homogeneous.

23. The cell population of claim 21 wherein said cell population is heterogeneous.

24. The cell population of claim 23 further comprising at least one cell type of bone marrow cells, chondrocytes, chondroblasts, chondrocyte progenitor cells, or stem cells.

25. A cell population comprising the postpartum-derived cell of claim 12.

26. The cell population of claim 25 wherein said cell population is substantially homogeneous.

27. The cell population of claim 25 wherein said cell population is heterogeneous.

28. The cell population of claim 27 further comprising at least one cell type of bone marrow cells, chondrocytes, chondroblasts, chondrocyte progenitor cells, stem cells, or other pluripotent or multipotent cell.

29. A cell population comprising the postpartum-derived cell of claim 20.

30. The cell population of claim 29 wherein said cell population is substantially homogeneous.

31. The cell population of claim 29 wherein said cell population is heterogeneous.

32. The cell population of claim 31 further comprising at least one cell type of bone marrow cells, osteocytes, osteoblasts, osteoclasts, bone lining cells, stem cells or other pluripotent or multipotent cell.

33. A cell lysate prepared from the cell population of claim 21.

34. A soluble cell fraction prepared from the cell lysate of claim 33.

35. A cell lysate prepared from the cell population of claim 25.

36. A soluble cell fraction prepared from the cell lysate of claim 35.

37. A cell lysate prepared from the cell population of claim 29.

38. A soluble cell fraction prepared from the cell lysate of claim 37.

39. An extracellular matrix of the cell population of claim 21.

40. An extracellular matrix of the cell population of claim 25.

41. An extracellular matrix of the cell population of claim 29.

42. A composition comprising the cell population of claim 21 and one or more bioactive factors.

43. A composition comprising the cell population of claim 25 and one or more bioactive factors.

44. A composition comprising the cell population of claim 29 and one or more bioactive factors.

45. The composition of claim 42 wherein said bioactive factor is a chondrogenic differentiation-inducing factor.

46. The composition of claim 42 wherein said bioactive factor is an osteogenic differentiation-inducing factor.

47. A pharmaceutical composition comprising a cell of claim 1 and a pharmaceutically acceptable carrier.

48. A pharmaceutical composition comprising a cell of claim 11 and a pharmaceutically acceptable carrier.

49. A pharmaceutical composition comprising a cell of claim 20 and a pharmaceutically acceptable carrier.

50. A pharmaceutical composition comprising the extracellular matrix of claim 39 and a pharmaceutically acceptable carrier.

51. A pharmaceutical composition comprising the extracellular matrix of claim 40 and a pharmaceutically acceptable carrier.

52. A pharmaceutical composition comprising the extracellular matrix of claim 41 and a pharmaceutically acceptable carrier.

53. A pharmaceutical composition comprising the lysate of claim 33 and a pharmaceutically acceptable carrier.

54. A pharmaceutical composition comprising the lysate of claim 35 and a pharmaceutically acceptable carrier.

55. A pharmaceutical composition comprising the lysate of claim 37 and a pharmaceutically acceptable carrier.

56. A cell culture comprising at least one cell of claim 1 in a culture medium.

57. The cell culture of claim 56 wherein said culture medium comprises chondrogenic medium or osteogenic medium.

58. The cell culture of claim 56 further comprising at least one chondrogenic differentiation-inducing agent.

59. The cell culture of claim 58 wherein said chondrogenic differentiation-inducing agent is at least one of transforming growth factor-beta3 or growth and differentiation factor-5.

60. The cell culture of claim 56 further comprising at least one osteogenic differentiation-inducing agent.

61. The cell culture of claim 60 wherein said osteogenic differentiation-inducing agent is at least one of transforming growth factor-beta 1, bone morphogenic protein (BMP)-2, or BMP4.

62. A matrix comprising a cell population of claim 21.

63. A matrix comprising a cell population of claim 25.

64. A matrix comprising a cell population of claim 29.

65. The matrix of claim 62 wherein said matrix comprises a three-dimensional scaffold.

66. The matrix of claim 63 wherein said matrix comprises a three-dimensional scaffold.

67. The matrix of claim 64 wherein said matrix comprises a three-dimensional scaffold.

68. A method of treating a condition in a patient comprising administering to said patient one or more postpartum-derived cells of claim 1.

69. The method of claim 68 wherein said condition is a bone or cartilage condition.

70. The method of claim 69 wherein said bone or cartilage condition is a congenital bone or cartilage defect, meniscal injury or defect, bone/spinal deformation, osteosarcoma, myeloma, bone dysplasia or scoliosis, osteoporosis, periodontal disease, dental bone loss, osteomalacia, rickets, fibrous osteitis, renal bone dystrophy, spinal fusion, spinal disc reconstruction or removal, Paget'"'"'s disease of bone, rheumatoid arthritis, osteoarthritis, or a traumatic or surgical injury.

71. The method of claim 68 wherein said postpartum-derived cells are administered with at least one other cell type.

72. The method of claim 71 wherein said at least one other cell type is bone marrow cells, chondrocytes, chondroblasts, chondrocyte progenitor cells, osteocytes, osteoblasts, osteoclasts, bone lining cells, stem cells, or other pluripotent or multipotent cell.

73. The method of claim 68 wherein said postpartum-derived cells are inoculated on a matrix.

74. The method of claim 73 wherein said matrix is implanted into said patient.

75. The method of claim 68 wherein said postpartum-derived cells are induced to differentiate to a chondrogenic or osteogenic phenotype prior to said step of administering.

76. The method of claim 68 wherein said postpartum-derived cells are co-administered with at least one bioactive factor.

77. The method of claim 68 wherein said cells are administered to a bone of said patient.

78. The method of claim 68 wherein said cells are administered to cartilage of said patient.

79. A method of regenerating a tissue in a patient in need thereof comprising administering the cell population of claim 21 to said patient.

80. The method of claim 79 wherein said tissue is bone or cartilage.

81. A method of regenerating a tissue in a patient in need thereof comprising administering the cell population of claim 25 to said patient.

82. The method of claim 81 wherein said tissue is bone or cartilage.

83. A method of regenerating a tissue in a patient in need thereof comprising administering the cell population of claim 29 to said patient.

84. The method of claim 83 wherein said tissue is bone or cartilage.

85. The method of claim 68 wherein said cells are implanted into said patient.

86. A conditioned medium generated by the growth of the culture of claim 56.

87. A method for identifying a compound that stimulates chondrogenesis of a postpartum-derived cell comprising contacting a cell of claim 1 with said compound and monitoring said cell for a marker of chondrogenesis.

88. A method for identifying a compound that stimulates osteogenesis of a postpartum-derived cell comprising contacting a cell of claim 1 with said compound and monitoring said cell for a marker of osteogenesis.

89. A method for identifying a compound that is toxic to a postpartum-derived cell of claim 1 comprising contacting said cell with said compound and monitoring survival of said cell.

90. A kit comprising at least one cell of claim 1 and at least one additional component of a matrix, a hydrating agent, a cell culture substrate, a differentiation-inducing agent, and cell culture media.

91. The kit of claim 90 wherein said matrix is a three-dimensional scaffold.

92. The kit of claim 91 wherein said cell is seeded on said scaffold.

93. The kit of claim 90 wherein said differentiation-inducing agent is an osteogenic differentiation-inducing agent or a chondrogenic differentiation-inducing agent.

94. A method of treating a patient having a bone or cartilage condition comprising administering to said patient the extracellular matrix of a cell of claim 1.

95. A method of treating a patient having a bone or cartilage condition comprising administering to said patient the cell lysate of claim 33.

96. A method of treating a patient having a bone or cartilage condition comprising administering to said patient the conditioned medium of claim 86.

97. The pharmaceutical composition of claim 47 wherein said composition comprises an effective amount of said cells to treat a bone or cartilage condition.

98. The pharmaceutical composition of claim 48 wherein said composition comprises an effective amount of said cells to treat a bone or cartilage condition.

99. The pharmaceutical composition of claim 49 wherein said composition comprises an effective amount of said cells to treat a bone or cartilage condition.

100. The pharmaceutical composition of claim 50 wherein said composition comprises an effective amount of said extracellular matrix to treat a bone or cartilage condition.

101. The pharmaceutical composition of claim 51 wherein said composition comprises an effective amount of said extracellular matrix to treat a bone or cartilage condition.

102. The pharmaceutical composition of claim 52 wherein said composition comprises an effective amount of said extracellular matrix to treat a bone or cartilage condition.

103. The pharmaceutical composition of claim 53 wherein said composition comprises an effective amount of said lysate to treat a bone or cartilage condition.

104. The pharmaceutical composition of claim 54 wherein said composition comprises an effective amount of said lysate to treat a bone or cartilage condition.

105. The pharmaceutical composition of claim 55 wherein said composition comprises an effective amount of said lysate to treat a bone or cartilage condition.

106. The pharmaceutical composition of claim 47 further comprising at least one other cell type of stem cells, bone marrow cells, chondrocytes, chondroblasts, osteocytes, osteoblasts, osteoclasts, bone lining cells, and other bone or cartilage progenitor cells.

107. The pharmaceutical composition of claim 48 further comprising at least one other cell type of stem cells, bone marrow cells, chondrocytes, chondroblasts, osteocytes, osteoblasts, osteoclasts, bone lining cells, and other bone or cartilage progenitor cells.

108. The pharmaceutical composition of claim 49 further comprising at least one other cell type of stem cells, bone marrow cells, chondrocytes, chondroblasts, osteocytes, osteoblasts, osteoclasts, bone lining cells, and other bone or cartilage progenitor cells.