POSTPARTUM CELLS DERIVED FROM UMBILICAL CORD TISSUE, AND METHODS OF MAKING AND USING THE SAME

• US 20100210013A1
• Filed: 02/19/2009
• Published: 08/19/2010
• 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., Seyda, Agnieszka, Harris, Ian Ross, Yi, Chin-Feng, Messina, Darin J., Mistry, Sanjay, Gosiewska, Anna, Kihm, Anthony J.
• Granted Patent: US 9,498,501 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• 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 cell obtained from mammalian umbilical cord tissue substantially free of blood:

wherein said isolated cell self-renews and expands its numbers in culture;

and wherein said isolated cell does not express CD117.

2. The isolated cell of claim 1, wherein said isolated cell expresses one or more of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A, B, C.

3. The isolated cell of claim 2, wherein said isolated cell expresses each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, and HLA-A, B, C.

4. The isolated cell of claim 2, wherein said isolated cell does not express one or more of CD31, CD34, CD45, CD141, and HLA-DR, DP, DQ.

5. The isolated cell of claim 1, wherein said isolated cell does not express any of CD31, CD34, CD45, CD141, and HLA-DR, DP, DQ.

6. The isolated cell of claim 3, wherein said isolated cell does not express any of CD31, CD34, CD45, CD141, and HLA-DR, DP, DQ

7. The isolated cell of claim 1, wherein said isolated cell expresses a gene for one or more 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.

8. The isolated cell of claim 1, wherein said isolated cell expresses a gene for each 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.

9. The isolated cell of claim 1, wherein said isolated 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.

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

11. The isolated cell of claim 1, wherein said isolated cell does not secrete one or more of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb and VEGF.

12. The isolated cell of claim 1, wherein said isolated cell does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb and VEGF.

13. The isolated cell of claim 1, wherein said isolated cell requires L-valine for growth.

14. The isolated cell of claim 1, wherein said isolated cell expands in the presence of from about 5% to about 20% oxygen.

15. The isolated cell of claim 1, wherein said isolated cell maintains a normal karyotype upon passaging.

16. The isolated cell of claim 1, wherein said isolated cell can undergo at least about twenty-five doublings in culture.

17. The isolated cell of claim 1, wherein said isolated cell can undergo at least about thirty doublings in culture.

18. The isolated cell of claim 1, wherein said isolated cell can undergo at least about forty doublings in culture.

19. The isolated cell of claim 1, wherein said isolated cell doubles at a rate sufficient to generate yields of greater than about 10¹⁴ cells in less than about 80 days in culture when seeded at about 10³ cells/cm².

20. The isolated cell of claim 1, wherein said isolated cell doubles at a rate sufficient to generate greater than about 10¹⁵ cells in less than about 80 days in culture when seeded at about 5×

10³ cells/cm².

21. The isolated cell of claim 1, wherein said isolated cell doubles at a rate sufficient to generate greater than about 10¹⁷ cells in less than about 65 days in culture when seeded at about 5×

10³ cells/cm².

22. The isolated cell of claim 21, wherein said doubling occurs in the presence of from about 5% to about 20% oxygen.

23. The isolated cell of claim 1, wherein said isolated cell is obtained from human umbilical cord tissue.

24. The isolated cell of claim 1, wherein said isolated cell is adherent.

25. The isolated cell of claim 1, wherein said isolated cell can differentiate into a cell of a different phenotype.

26. A substantially homogeneous cell population comprising the isolated cell of claim 1.

27. A cell culture comprising the isolated cell of claim 1.

28. The cell culture of claim 27, further comprising L-valine.

29. A conditioned medium made by culturing the isolated cell of claim 1.

30. The conditioned medium of claim 29, comprising one or more of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

31. A three dimensional matrix comprising the isolated cell of claim 1.

32. The three-dimensional matrix of claim 31, wherein said matrix comprises a biocompatible or bioabsorbable polymer.

33. The three-dimensional matrix of claim 32, wherein said matrix comprises a nonwoven mat comprising a copolymer of glycolic and lactic acids, PCL/PGA copolymer, or self assembling peptides.

34. An isolated cell obtained from mammalian umbilical cord tissue substantially free of blood;

wherein said isolated cell self-renews and can undergo at least about thirty doublings in culture; and

wherein said isolated cell requires L-valine for growth.

35. The isolated cell of claim 34, wherein said isolated cell expands in the presence of from about 5% to about 20% oxygen.

36. The isolated cell of claim 34, wherein said isolated cell maintains a normal karyotype upon passaging.

37. The isolated cell of claim 34, wherein said isolated cell doubles at a rate sufficient to generate yields of greater than about 10¹⁴ cells in less than about 80 days in culture when seeded at about 10³ cells/cm².

38. The isolated cell of claim 34, wherein said isolated cell doubles at a rate sufficient to generate greater than about 10¹⁵ cells in less than about 80 days in culture when seeded at about 5×

10³ cells/cm².

39. The isolated cell of claim 34, wherein said isolated cell doubles at a rate sufficient to generate greater than about 10¹⁷ cells in less than about 65 days in culture when seeded at about 5×

10³ cells/cm².

40. The isolated cell of claim 39, wherein said doubling occurs in the presence of from about 5% to about 20% oxygen.

41. The isolated cell of claim 34, wherein said isolated cell is obtained from human umbilical cord tissue.

42. The isolated cell of claim 34, wherein said isolated cell is adherent.

43. The isolated cell of claim 34, wherein said isolated cell expresses a gene for one or more 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.

44. The isolated cell of claim 34, wherein said isolated cell expresses a gene for each 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.

45. The isolated cell of claim 34, wherein said isolated 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.

46. The isolated cell of claim 34, wherein said isolated cell secretes each of the factors MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

47. The isolated cell of claim 45, wherein said isolated cell does not secrete one or more of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb and VEGF.

48. The isolated cell of claim 46, wherein said isolated cell does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb and VEGF.

49. The isolated cell of claim 34, wherein said isolated cell can differentiate into a cell of a different phenotype.

50. A substantially homogeneous cell population comprising the isolated cell of claim 34.

51. A cell culture comprising the isolated cell of claim 34.

52. The cell culture of claim 51, further comprising L-valine.

53. A conditioned medium made by culturing the isolated cell of claim 34.

54. The conditioned medium of claim 53, comprising one or more of MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, and TIMP1.

55. A three dimensional matrix comprising the isolated cell of claim 34.

56. The three-dimensional matrix of claim 55, wherein said matrix comprises a biocompatible or bioabsorbable polymer.

57. The three-dimensional matrix of claim 56, wherein said matrix comprises a nonwoven mat comprising a copolymer of glycolic and lactic acids, PCL/PGA copolymer, or self assembling peptides.

58. The isolated cell of claim 34, wherein said isolated cell can undergo at least about forty doublings in culture.

59. The isolated cell identified by ATCC Accession No. PTA-6067.

60. The isolated cell identified by ATCC Accession No. PTA-6068.