Postpartum cells derived from umbilical cord tissue, and methods of making and using the same

• US 20050054098A1
• Filed: 06/25/2004
• Published: 03/10/2005
• Est. Priority Date: 06/27/2003
• Status: Active Grant

• Current Assignee: Depuy Synthes Products Incorporated (Johnson & Johnson)
• Original Assignee: Ethicon Incorporated (Johnson & Johnson)
• Inventors: Harmon, Alexander M., Messina, Darin J., Mistry, Sanjay, Gosiewska, Anna, Seyda, Agnieszka, Kihm, Anthony J., Harris, Ian Ross, Yi, Chin-Feng
• Granted Patent: US 7,510,873 B2
• Time in Patent Office: Days
• US Class Current: 435/372
• 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 human umbilical cords are disclosed along with methods for their therapeutic use. Isolation techniques, culture methods and detailed characterization of the cells with respect to their cell surface markers, gene expression, and their secretion of trophic factors are described.

Claims

1. An isolated umbilicus-derived cell comprising a cell derived from mammalian umbilical cord tissue substantially free of blood, said cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes.

2. The isolated cell of claim 1 which can expand in the presence of oxygen from about 5% to about 20%.

15. A cell culture comprising the isolated cell of claim 1 which is free of maternal cells.

16. The isolated cell of claim 1 which maintains a normal karyotype as it is passaged.

17. The isolated cell of claim 1 wherein the cell is characterized in its production or lack of production of one or more cell surface markers comprising CD10, CD13, CD31, CD44, CD45, CD73, CD90, CD117, CD141, PDGFr-alpha, HLA-A,B,C, and HL-Dr, DP,DQ.

18. The isolated cell of claim 17 wherein the cell produces one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, or HLA-A,B,C.

19. The isolated cell of claim 18 wherein the cell produces each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A,B,C.

20. The isolated cell of claim 17 wherein the cell does not produce one or more of CD31, CD34, CD45, CD117, CD141, or HLA-DR,DP,DQ, as detected by flow cytometry.

21. The isolated cell of claim 20 wherein the cell produces one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, or HLA-A,B,C.

22. The isolated cell of claim 20 wherein the cell does not produce any of CD31, CD34, CD45, CD117, CD141, or HLA-DR,DP,DQ, as detected by flow cytometry.

23. The isolated cell of claim 22 wherein the cell produces each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A,B,C.

83. A conditioned culture medium generated by the growth of the culture of claim 15 or 82.

84. The conditioned medium of claim 83 comprising one or more of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.

85. A mammalian cell culture comprising the conditioned medium of claim 84 and a mammalian cell in need of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.

86. The culture of claim 15 or claim 82 comprising the umbilicus-derived cells and another mammalian cell of any phenotype.

87. The culture of claim of claim 86 comprising a human cell line in addition to the umbilicus-derived cell.

88. A three dimensional matrix comprising the umbilicus-derived cell of claim 1.

89. The three-dimensional matrix of claim 88 wherein the matrix comprises a biocompatible or bioabsorbable polymer.

90. The three-dimensional matrix of claim 89 wherein the matrix comprises Vicryl®

matrix, PCL/PGA copolymer, or self assembling peptides.

91. An implantable tissue structure comprising the matrix of claim 89.

3. The cell of claim

4. The isolated cell of claim 3 which can double sufficiently to generate yields of greater than about 10¹⁴ cells in less than about 80 days in culture when seeded at about 10³ cells/cm².

5. The isolated cell of claim 3 which can double sufficiently to generate greater than about 10¹⁵ cells in less than about 80 days in culture when seeded at about 5×

10³ cells/cm².

6. The isolated cell of claim 5 which can double sufficiently to generate greater than about 10¹⁷ cells in less than about 65 days in culture when seeded at about 5×

10³ cells/cm².

7. The isolated cell of claim 3 which can undergo at least 40 doublings in culture.

8. The cell of claim 3 which is isolated from human umbilicus.

9. The isolated cell of claim 8 isolated in the presence of one or more enzyme activities comprising metalloproteases, neutral proteases, or mucolytic enzymes.

10. The isolated cell of claim 9 wherein the enzyme activities include at least one collagenase, and one or more of the protease activities, dispase and thermolysin.

11. The isolated cell of claim 9 wherein the enzyme activities are a collagenase from Clostridium histolyticum and dispase.

12. The isolated cell of claim 11 wherein the enzyme activities further include hyaluronidase.

13. The isolated cell of claim 12 which attaches and expands on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating with gelatin, laminin, collagen, polyornithine, polylysine, vitronectin, or fibronectin.

14. The isolated cell of claim 13 which expands in the presence of from about 2% to about 15% added serum, in the presence or absence of beta-mercaptoethanol, and in the presence or absence of added growth factors including EGF, FGF, PDGF, VEGF, IGF or LIF.

94. A human tissue comprising the cell of claim 8.

95. A cell lysate derived from the cell of claim 8.

96. A soluble cell fraction derived from the cell of claim 8

97. A membrane-enriched cell fraction derived from the cell of claim 8.

98. An extracellular membrane fraction derived from the cell of claim 8.

24. An isolated CD45⁻

umbilicus-derived cell having a cell surface marker profile wherein the cell produces one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, or HLA-A,B,C, and does not express one or more of CD31, CD34, CD117, CD141, or HLA-DR,DP,DQ, as detected by flow cytometry.

25. The isolated cell of claim 24 wherein the cell surface marker expression profile remains substantially unchanged with passage, culture vessel surface coating, or isolation procedure.

26. The isolated cell of claim 25 expressing a gene for one or more of interleukin 8;

reticulon 1;

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

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

     2);

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

or tumor necrosis factor, alpha-induced protein 3.

27. The isolated cell of claim 26 expressing a gene for each of interleukin 8;

reticulon 1;

chemokine (C-X-C motif) ligand 1 (melonoma 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.

28. The isolated cell of claim 27 wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, an ileac crest bone marrow cell, or placenta-derived cell.

29. The isolated cell of claim 25, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, has reduced expression of one or more genes selected from the group consisting of:

short stature homeobox 2;

heat shock 27 kDa protein 2;

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

     1);

elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);

Homo sapiens mRNA;

cDNA DKFZp586M2022 (from clone DKFZp586M2022);

mesenchyme homeobox 2 (growth arrest-specific homeobox);

sine oculis homeobox homolog 1 (Drosophila);

crystallin, alpha B;

dishevelled associated activator of morphogenesis 2;

DKFZP586B2420 protein;

similar to neuralin 1;

tetranectin (plasminogen binding protein);

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 (Drosophila);

hypothetical gene BC008967;

collagen, type VIII, alpha 1;

tenascin C (hexabrachion);

iroquois homeobox protein 5;

hephaestin;

integrin, beta 8;

synaptic vesicle glycoprotein 2;

Homo sapiens 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 (Drosophila);

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);

Homo sapiens mRNA full length insert cDNA clone EUROIMAGE 1968422;

EphA3;

KIAA0367 protein;

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

hypothetical protein FLJ14054;

Homo sapiens mRNA;

cDNA DKFZp564B222 (from clone DKFZp564B222);

vesicle-associated membrane protein 5 (myobrevin);

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.

30. The isolated cell of claim 29 expressing a gene for one or more of interleukin 8;

reticulon 1;

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

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

     2);

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

or tumor necrosis factor, alpha-induced protein 3, wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, an ileac crest bone marrow cell, or placenta-derived cell.

31. The isolated cell of claim 29, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, has reduced expression of genes for each of:

short stature homeobox 2;

heat shock 27 kDa protein 2;

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

     1);

elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);

Homo sapiens mRNA;

cDNA DKFZp586M2022 (from clone DKFZp586M2022);

mesenchyme homeobox 2 (growth arrest-specific homeobox);

sine oculis homeobox homolog 1 (Drosophila);

crystallin, alpha B;

dishevelled associated activator of morphogenesis 2;

DKFZP586B2420 protein;

similar to neuralin 1;

tetranectin (plasminogen binding protein);

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 (Drosophila);

hypothetical gene BC008967;

collagen, type VIII, alpha 1;

tenascin C (hexabrachion);

iroquois homeobox protein 5;

hephaestin;

integrin, beta 8;

synaptic vesicle glycoprotein 2;

Homo sapiens 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 (Drosophila);

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);

Homo sapiens mRNA full length insert cDNA clone EUROIMAGE 1968422;

EphA3;

KIAA0367 protein;

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

hypothetical protein FLJ14054;

Homo sapiens mRNA;

cDNA DKFZp564B222 (from clone DKFZp564B222);

vesicle-associated membrane protein 5 (myobrevin);

EGF-containing fibulin-like extracellular matrix protein 1;

BCL2/adenovirus EIB 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.

32. The isolated cell of claim 31 expressing a gene for each of interleukin 8;

reticulon 1;

chemokine (C-X-C motif) ligand 1 (melonoma 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, wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell.

33. An isolated human umbilicus-derived cell, which relative to a human cell that is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell, has reduced expression of genes for each of:

short stature homeobox 2;

heat shock 27 kDa protein 2;

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

     1);

elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);

Homo sapiens mRNA;

cDNA DKFZp586M2022 (from clone DKFZp586M2022);

mesenchyme homeobox 2 (growth arrest-specific homeobox);

sine oculis homeobox homolog 1 (Drosophila);

crystallin, alpha B;

dishevelled associated activator of morphogenesis 2;

DKFZP586B2420 protein;

similar to neuralin 1;

tetranectin (plasminogen binding protein);

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 (Drosophila);

hypothetical gene BC008967;

collagen, type VIII, alpha 1;

tenascin C (hexabrachion);

iroquois homeobox protein 5;

hephaestin;

integrin, beta 8;

synaptic vesicle glycoprotein 2;

Homo sapiens 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 (Drosophila);

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);

Homo sapiens mRNA full length insert cDNA clone EUROIMAGE 1968422;

EphA3;

KIAA0367 protein;

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

hypothetical protein FLJ14054;

Homo sapiens mRNA;

cDNA DKFZp564B222 (from clone DKFZp564B222);

vesicle-associated membrane protein 5 (myobrevin);

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;

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

which expresses a gene for each of interleukin 8;

reticulon 1;

chemokine (C-X-C motif) ligand 1 (melonoma 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, wherein the expression is increased relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, or an ileac crest bone marrow cell.

34. The isolated cell of claim 33 capable of self-renewal and expansion in culture, and having the potential to differentiate into cells of other phenotypes.

35. The isolated cell of claim 34 which produces one or both of vimentin and alpha smooth muscle actin.

36. The isolated cell of claim 35 which produces both vimentin and alpha-smooth muscle actin.

37. The isolated cell of claim 36 wherein the production of vimentin and alpha smooth muscle actin is retained over passaging under growth conditions.

38. A therapeutic cell culture comprising the cell of claim 34.

39. The therapeutic cell culture of claim 38 which does not substantially stimulate allogeneic PBMCs.

40. The therapeutic cell culture of claim 39 which lacks detectable amounts of HLA-DR, HLA-DP, HLA-DQ, CD80, CD86, and B7-H2, as determined by flow cytometry.

41. The therapeutic cell culture of claim 40 which further lacks detectable amounts of HLA-G and CD178, as determined by flow cytometry.

42. The therapeutic cell culture of claim 41, which produces detectable amounts of PD-L2, as determined by flow cytometry.

43. The therapeutic cell culture of claim 34 which does not substantially stimulate a lymphocyte mediated response in vitro, as compared to allogeneic controls in a mixed lymphocyte reaction.

44. An isolated human umbilicus-derived cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes, wherein the cell does not substantially stimulate naï

ve CD4⁺ T cells, and expresses PD-L2, but not HLA-G, CD178, HLA-DR, HLA-DP, HLA-DQ, CD80, CD86, or B7-H2.

45. The isolated cell of claim 44 wherein the cell produces vimentin and alpha-smooth muscle actin.

46. The isolated cell of claim 44 wherein the cell secretes one or more cellular factors.

47. The isolated cell of claim 46 wherein the factors are MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.

48. The isolated cell of claim 47 wherein the cell secretes each of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

49. The isolated cell of claim 44 wherein the cell does not secrete one or more of the cellular factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.

50. The isolated cell of claim 49, wherein the cell secretes one or more of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

51. The isolated cell of claim 50 wherein the cell secretes each of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

52. The isolated cell of claim 49 wherein the cell does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.

53. The isolated cell of claim 52, wherein the cell secretes one or more of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1.

54. The isolated cell of claim 53 wherein the cell secretes each of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

57. The isolated cell of claim 50 wherein the cell does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.

99. An injectable therapeutic cell comprising an isolated human umbilicus-derived cell of claim 44.

100. The injectable cell of claim 99 treated to inactivate tissue factor.

101. The injectable cell of claim 100 wherein the treated is with an anti-tissue factor antibody.

55. An isolated human umbilicus-derived cell which secretes one or more of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, or TIMP1, and does not secrete one or more of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb or VEGF, as detected by ELISA.

56. The isolated cell of claim 55 which secretes each of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

58. A therapeutic cell culture comprising the isolated cell of claim 55.

59. The therapeutic cell culture of claim 58 further comprising one or more of a pharmaceutically-acceptable carrier, another cell culture, an antiapoptotic compound, an antithrombogenic compound, an anti-inflammatory compound, an immunosuppressive compound, an immunomodulatory compound, an angiogenic factor, and a neurotrophic factor.

60. A method of deriving, from umbilical tissue, an isolated cell, said cell capable of self-renewal and expansion in culture, and having the potential to differentiate into cells of other phenotypes, the method comprising the steps of:

(a) obtaining umbilical tissue;

(b) removing substantially all of blood to yield a substantially blood-free umbilical tissue,

(c) dissociating the tissue by mechanical or enzymatic treatment, or both,

(d) resuspending the tissue in a culture medium, and

(e) providing growth conditions which allow for the growth of an umbilicus-derived cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes.

61. The method of claim 60 further comprising the step of selecting adherent cells after from about ten to about 100 hours in culture.

62. The method of claim 61 wherein the umbilicus tissue is from a human.

63. The method of claim 62 wherein the derived cell is capable of to expansion in the absence of added growth factors in medium with from 2% to about 15% added serum.

64. The method of claim 63 wherein the derived cell can expand in the presence of oxygen from about 5% to about 20%.

65. The method of claim 64 wherein the derived cell cannot be maintained in the absence of L-valine.

66. The method of claim 65 wherein the derived cell can undergo at least 40 doublings in culture.

67. The method of claim 65 wherein the derived cell can double sufficiently to generate at least about 10¹⁷ cells in less than about 65 days in culture when seeded at about 5×

10³ cells/cm².

68. The method of claim 62 wherein the umbilical tissue is obtained after normal or surgically-assisted child birth from a full-term or pre-term pregnancy.

69. The method of claim 60 wherein the dissociation step comprises the use of one or more enzyme activities selected from the group consisting of metalloprotease, hyaluronidase, and neutral protease.

70. The method of claim 69 wherein the enzyme activities are collagenase and dispase.

71. The method of claim 70 wherein the enzyme activities further include hyaluronidase.

72. The method of claim 71 wherein the dissociating step comprises incubating at about 37°

C.

73. The method of claim 72 wherein the incubating is for one or more hours.

74. The method of claim 72 where the incubating is for about 2 hours.

75. The method of claim 69 wherein the derived cell attaches and expands on a coated or uncoated tissue culture vessel, wherein the coated tissue culture vessel comprises a coating with gelatin, laminin, collagen, polyomithine, polylysine, vitronectin, or fibronectin.

76. The method of claim 69 wherein the derived cell expands in the presence of from about 2% to about 15% Fetal Bovine Serum, in the presence or absence of beta-mercaptoethanol, and in the presence or absence of one or more added growth factors including EGF, FGF, PDGF, VEGF, IGF and LIF.

77. The method of claim 60 wherein the removing step comprises removal of free or clotted blood by one or more of washing, suctioning, blotting, centrifugal separation, or enzymatic removal.

78. The method of claim 60 wherein the dissociating step is accomplished aseptically.

79. The method of claim 78 wherein the dissociation step comprises one or more of mincing, blending, homogenizing, or grinding.

80. An isolated human umbilicus-derived cell derived by the method of claim 69.

81. The isolated cell of claim 80 which maintains a normal karyotype with passaging.

82. A therapeutic culture of human umbilicus-derived cells derived by the method of claim 69, wherein the culture is free of maternal cells.

92. An implantable device comprising the therapeutic cell of claim 82.

93. An implantable human tissue matrix comprising the cell of claim 80.

102. An isolated postpartum-derived cell comprising an L-valine-requiring cell derived from human postpartum tissue substantially free of blood, said cell capable of self-renewal and expansion in culture and having the potential to differentiate into a cell of cardiomyocyte phenotypes;

the cell capable of growth in an atmosphere containing oxygen from about 5% to at least about 20%;

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

potential for at least about 40 doublings in culture;

attachment and expansion on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyornithine, vitronectin, or fibronectin;

production of at least one of tissue factor, vimentin, and alpha-smooth muscle actin;

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

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;

expression of 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;

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 (supravalvular aortic stenosis, Williams-Beuren syndrome);

Homo sapiens mRNA;

cDNA DKFZp586M2022 (from clone DKFZp586M2022);

mesenchyme homeobox 2 (growth arrest-specific homeobox);

sine oculis homeobox homolog 1 (Drosophila);

crystallin, alpha B;

dishevelled associated activator of morphogenesis 2;

DKFZP586B2420 protein;

similar to neuralin 1;

tetranectin (plasminogen binding protein);

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 FLJ23 191;

interleukin 11 receptor, alpha;

procollagen C-endopeptidase enhancer;

frizzled homolog 7 (Drosophila);

hypothetical gene BC008967;

collagen, type VIII, alpha 1;

tenascin C (hexabrachion);

iroquois homeobox protein 5;

hephaestin;

integrin, beta 8;

synaptic vesicle glycoprotein 2;

Homo sapiens cDNA FLJ12280 fis, clone MAMMA 1001744;

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 (Drosophila);

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);

Homo sapiens mRNA full length insert cDNA clone EUROIMAGE 1968422;

EphA3;

KLAA0367 protein;

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

hypothetical protein FLJ14054;

Homo sapiens mRNA;

cDNA DKFZp564B222 (from clone DKFZp564B222);

vesicle-associated membrane protein 5 (myobrevin);

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;

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

secretion of at least one of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1a, RANTES, and TIMP1; and

lack of secretion of at least one of TGF-beta2, ANG2, PDGFbb, MIP1beta, I309, MDC, and VEGF, as detected by ELISA.

103. The isolated postpartum-derived cell of claim 102;

wherein said cell comprises each of the following characteristics;

potential for at least about 40 doublings in culture;

attachment and expansion on a coated or uncoated tissue culture vessel, wherein a coated tissue culture vessel comprises a coating of gelatin, laminin, collagen, polyomithine, vitronectin, or fibronectin;

production of tissue factor, vimentin, and alpha-smooth muscle actin;

production of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2 and HLA-A,B,C;

lack of production of each of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, and HLA-DR,DP,DQ, as detected by flow cytometry;

expression 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;

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 each of;

short stature homeobox 2;

heat shock 27 kDa protein 2;

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

     1);

elastin (supravalvular aortic stenosis, Williams-Beuren syndrome);

Homo sapiens mRNA;

cDNA DKFZp586M2022 (from clone DKFZp586M2022);

mesenchyme homeobox 2 (growth arrest-specific homeobox);

sine oculis homeobox homolog 1 (Drosophila);

crystallin, alpha B;

dishevelled associated activator of morphogenesis 2;

DKFZP586B2420 protein;

similar to neuralin 1;

tetranectin (plasminogen binding protein);

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 (Drosophila);

hypothetical gene BC008967;

collagen, type VIII, alpha 1;

tenascin C (hexabrachion);

iroquois homeobox protein 5;

hephaestin;

integrin, beta 8;

synaptic vesicle glycoprotein 2;

Homo sapiens cDNA FLJ12280 fis, clone MAMMA 1001744;

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 (Drosophila);

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);

Homo sapiens mRNA full length insert cDNA clone EUROIMAGE 1968422;

EphA3;

KIAA0367 protein;

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

hypothetical protein FLJ 4054;

Homo sapiens mRNA;

cDNA DKFZp564B222 (from clone DKFZp564B222);

vesicle-associated membrane protein 5 (myobrevin);

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;

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

secretion of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1a, RANTES, and TIMP1; and

lack of secretion of each of TGF-beta2, ANG2, PDGFbb, MIP1beta, I309, MDC, and VEGF, as detected by ELISA.