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

• US 10,500,234 B2
• Filed: 07/27/2017
• Issued: 12/10/2019
• 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: Mistry, Sanjay, Kihm, Anthony J., Harris, Ian R., Harmon, Alexander M., Messina, Darin J., Seyda, Agnieszka, Yi, Chin-Feng, Gosiewska, Anna
• Primary Examiner(s): Kim, Taeyoon
• Application Number:
  US15/661,878
• Publication Number:
  US 20170326183A1
• Time in Patent Office: 866 Days
• 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. A cell culture comprising a culture medium and an isolated cell derived from human umbilical cord tissue substantially free of blood, wherein said isolated cell is capable of self-renewal and expansion in culture, has the potential to differentiate into cells of other phenotypes, produces granulocyte chemotactic protein-2 (GCP-2), and can double sufficiently to generate yields of greater than about 1×

10¹⁷ cells in about 60 days in culture when seeded at about 5,000 cells/cm² in a flask or yields of greater than about 1×

10¹¹ cells in about 60 days in culture when seeded at about 1,000 cells/cm² in a flask.

2. The cell culture of claim 1, wherein the isolated cell can double sufficiently to generate greater than about 1×

10¹⁷ cells in about 60 days in culture when seeded at about 5,000 cells/cm² in a flask.

3. The cell culture of claim 1, wherein the isolated cell can double sufficiently to generate greater than about 1×

10¹¹ cells in about 60 days in culture when seeded at about 1,000 cells/cm² in a flask.

4. The cell culture of claim 1, wherein the isolated cell expresses CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, HLA-A, HLA-B, and HLA-C, does not express CD31, CD34, CD45, CD117, CD141, HLA-DR, HLA-DP and HLA-DQ, and has increased expression, relative to a human fibroblast, mesenchymal stem cell, or iliac crest bone marrow cell, of a gene for interleukin 8, reticulon 1 and chemokine (C-X-C motif) ligand 3.

5. The cell culture of claim 1, wherein the culture medium is a chemically-defined medium or a medium with added mammalian serum.

6. The cell culture of claim 5, wherein the culture medium comprises L-valine.

7. The cell culture of claim 1, wherein the isolated cell secretes MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1b, RANTES, and TIMP1, and does not secrete SDF-1alpha, TGF-beta2, ANG2, PDGFbb, MIP1a, and VEGF.

8. The cell culture of claim 1, wherein the isolated cell is cryopreserved.

9. A kit comprising the cell culture of claim 1 and at least one other component selected from the group consisting of a scaffold, a hydrating agent and a cell culture substrate.

10. A three-dimensional matrix comprising an isolated cell derived from human umbilical cord tissue substantially free of blood, wherein said isolated cell is capable of self-renewal and expansion in culture, has the potential to differentiate into cells of other phenotypes, produces granulocyte chemotactic protein-2 (GCP-2), and can double sufficiently to generate yields of greater than about 1×

10¹⁷ cells in about 60 days in culture when seeded at about 5,000 cells/cm² in a flask or yields of greater than about 1×

10¹¹ cells in about 60 days in culture when seeded at about 1,000 cells/cm² in a flask.

11. The three-dimensional matrix of claim 10, wherein the isolated cell expresses CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, HLA-A, HLA-B, and HLA-C, does not express CD31, CD34, CD45, CD117, CD141, HLA-DR, HLA-DP and HLA-DQ, and has increased expression, relative to a human fibroblast, mesenchymal stem cell, or iliac crest bone marrow cell, of a gene for interleukin 8, reticulon 1 and chemokine (C-X-C motif) ligand 3.

12. The three-dimensional matrix of claim 10, wherein the isolated cell secretes MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1b, RANTES, and TIMP1, and does not secrete SDF-1alpha, TGF-beta2, ANG2, PDGFbb, MIP1a, and VEGF.

13. The three-dimensional matrix of claim 10, wherein said matrix comprises a synthetic biocompatible or bioabsorbable polymer.

14. The three-dimensional matrix of claim 13, wherein said matrix comprises a nonwoven mat comprising a synthetic copolymer of glycolic and lactic acids, a PCL/PGA copolymer, or self-assembling peptides.

15. A cryopreserved composition comprising an isolated cell derived from human umbilical cord tissue substantially free of blood, wherein said isolated cell is capable of self-renewal and expansion in culture, has the potential to differentiate into cells of other phenotypes, produces granulocyte chemotactic protein-2 (GCP-2), and can double sufficiently to generate yields of greater than about 1×

10¹⁷ cells in about 60 days in culture when seeded at about 5,000 cells/cm² in a flask or yields of greater than about 1×

10¹¹ cells in about 60 days in culture when seeded at about 1,000 cells/cm² in a flask.

16. The cryopreserved composition of claim 15, wherein the isolated cell can double sufficiently to generate greater than about 1×

10¹⁷ cells in about 60 days in culture when seeded at about 5,000 cells/cm² in a flask.

17. The cryopreserved composition of claim 15, wherein the isolated cell can double sufficiently to generate greater than about 1×

10¹¹ cells in about 60 days in culture when seeded at about 1,000 cells/cm² in a flask.

18. The cryopreserved composition of claim 15, wherein the isolated cell expresses CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, HLA-A, HLA-B, and HLA-C, does not express CD31, CD34, CD45, CD117, CD141, HLA-DR, HLA-DP and HLA-DQ, and has increased expression, relative to a human fibroblast, mesenchymal stem cell, or iliac crest bone marrow cell, of a gene for interleukin 8, reticulon 1 and chemokine (C-X-C motif) ligand 3.

19. The cryopreserved composition of claim 15, wherein the isolated cell secretes MCP-1, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, MIP1b, RANTES, and TIMP1, and does not secrete SDF-1alpha, TGF-beta2, ANG2, PDGFbb, MIP1a, and VEGF.