METHODS AND COMPOSITIONS FOR THE CLINICAL DERIVATION OF AN ALLOGENIC CELL AND THERAPEUTIC USES

• US 20130216505A1
• Filed: 12/31/2012
• Published: 08/22/2013
• Est. Priority Date: 12/30/2011
• Status: Active Grant

• Current Assignee: Jadi Cell LLC
• Original Assignee: Amit Patel
• Inventors: Patel, Amit
• Granted Patent: US 9,803,176 B2
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/10 for impotence
• 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 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue
• CPC Class Codes: C12N 5/0654 Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0655 Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0657 : Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0665 : Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
• CPC Class Codes: C12N 5/0682 : Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells oocytes C12N5/0609

Abstract

Various cells, stem cells, and stem cell components, including associated methods of generating and using such cells are provided. In one aspect, for example, an isolated cell that is capable of self-renewal and culture expansion and is obtained from a subepithelial layer of a mammalian umbilical cord tissue. Such an isolated cell expresses at least three cell markers selected from CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and does not express at least three cell markers selected from CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.

Claims

1. An isolated cell obtained from a subepithelial layer of a mammalian umbilical cord tissue capable of self-renewal and culture expansion;

wherein the isolated cell expresses at least three cell markers selected from the group consisting of CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105; and

wherein the isolated cell does not express at least three cell markers selected from the group consisting of CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.

2. The isolated cell of claim 1, wherein the isolated cell expresses CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, and CD105.

3. The isolated cell of claim 1, wherein the isolated cell does not express CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, and HLA-DR.

4. The isolated cell of claim 1, wherein the isolated cell is positive for SOX2.

5. The isolated cell of claim 1, wherein the isolated cell is positive for OCT4.

6. The isolated cell of claim 1, wherein the isolated cell is positive for SOX2 and OCT4.

7. The isolated cell of claim 1, wherein the wherein the isolated cell is capable of differentiation into a cell type selected from the group consisting of adipocytes, chondrocytes, osteocytes, cardiomyocytes, endothelial cells, and myocytes.

8. The isolated cell of claim 1, wherein the isolated cell produces exosomes expressing CD63, CD9, or CD63 and CD9.

9. The isolated cell of claim 1 that has been differentiated into an adipocyte cell.

10. The isolated cell of claim 1 that has been differentiated into a chondrocyte cell.

11. The isolated cell of claim 1 that has been differentiated into an osteocyte cell.

12. The isolated cell of claim 1 that has been differentiated into a cardiomyocyte cell.

13. The isolated cell of claim 1 that has been expanded into a cell culture.

14. An isolated exosome derived from the isolated cell of claim 1, wherein the exosome expresses CD63, CD9, or CD63 and CD9.

15. A culture of differentiated cells derived from the isolated cell of claim 1, wherein the culture of differentiated cells includes a cell type selected from the group consisting of adipocytes, chondrocytes, osteocytes, cardiomyocytes, endothelial cells, myocytes and combinations thereof.

29. A method of treating a medical condition responsive to treatment with cells as recited in claim 9, comprising introducing cells of claim 9 into an individual having the medical condition.

30. The method of claim 29, wherein the medical condition includes a member selected from the group consisting of COPD, diabetes, ischemia, osteoarthritis, orthopedic damage, liver damage, chronic refractory angina, erectile dysfunction, spinal cord injuries, herniated disks, congestive heart failure, asthma, emphysema, wounds, acute radiation syndrome, autoimmune disorders, ischemic organ beds, graft vs. host disease, and combinations thereof.

31. The method of claim 29, wherein introducing cells into an individual further includes retrograde or antegrade delivery of the cells into an organ of the individual.

32. The method of claim 31, wherein the organ includes a member selected from the group consisting of the heart, the liver, a kidney, the brain, pancreas, and combinations thereof.

33. A method of treating a medical condition responsive to treatment with the differentiated cells as recited in claim 15, comprising introducing at least one cell type of the differentiated cells of claim 15 into an individual having the medical condition.

34. The method of claim 33, wherein the medical condition includes a member selected from the group consisting of COPD, diabetes, ischemia, osteoarthritis, orthopedic damage, liver damage, chronic refractory angina, congestive heart failure, asthma, emphysema, wounds, acute radiation syndrome, erectile dysfunction, spinal cord injury, herniated disks, renal failure, neurological disorders, autoimmune disorders, and combinations thereof.

16. A method of culturing stem cells from a subepithelial layer of a mammalian umbilical cord, comprising:

dissecting the subepithelial layer from the umbilical cord;

placing the dissected subepithelial layer interior side down on a substrate such that an interior side of the subepithelial layer is in contact with the substrate;

culturing the subepithelial layer on the substrate; and

removing explants for primary cell expansion.

17. The method of claim 16, wherein dissecting the subepithelial layer further includes removing Wharton'"'"'s Jelly from the umbilical cord.

18. The method of claim 16, wherein the subepithelial layer is cultured in a culture medium comprising a platelet lysate.

19. The method of claim 18, wherein the platelet lysate is a human or animal platelet lysate.

20. The method of claim 18, wherein the culture media is derived from human-free and animal-free ingredients.

21. The method of claim 16, wherein the substrate is a culture dish.

22. The method of claim 21, wherein the culture dish is a cell culture treated plastic, and the subepithelial layer is placed thereon without any additional pretreatment to the cell culture treated plastic.

23. The method of claim 16, wherein the substrate is a semi-solid cell culture substrate.

24. The method of claim 16, wherein the subepithelial layer is cultured in a normoxic environment.

25. The method of claim 16, wherein the subepithelial layer is cultured in a hypoxic environment.

26. The method of claim 16, wherein the culturing of the subepithelial layer and the removal of the explants are performed without the use of any enzymes.

27. The method of claim 16, further comprising subculturing the explants.

28. The method of claim 27, wherein the subculturing is performed without the use of any enzymes.

35. A method of treating COPD, comprising:

administering a COPD effective active agent intravenously to a subject to deliver the COPD effective active agent to a lower half of the subject'"'"'s lung; and

administering the COPD effective active agent in an aerosolized form to the subject via ventilation to deliver the COPD effective active agent to an upper half of the subject'"'"'s lung.

36. The method of claim 35, wherein the COPD effective active agent includes stem cells.

37. The method of claim 36, wherein the stem cells include the cells of claim 9.

38. The method of claim 36, wherein the stem cells are aerosolized with an aerosolizer to a size of from about 6 to about 200 microns.

39. The method of claim 35, wherein the administration is concomitant.

40. The method of claim 35, wherein the administration is sequential.

41. The method of claim 35, wherein the COPD effective active agent includes a member selected from the group consisting of exosomes, cell lysates, protein extracts derived from cell culture, and combinations thereof.