Post-partum mammalian placenta, its use and placental stem cells therefrom

• US 20040048372A1
• Filed: 08/12/2003
• Published: 03/11/2004
• Est. Priority Date: 12/06/2000
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Hariri, Robert J.
• Granted Patent: US 7,255,879 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 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61M 1/02 Blood transfusion apparatus blood infusion by syringes A61M5/14
• CPC Class Codes: A61M 2202/0462 Placental blood, umbilical cord blood
• CPC Class Codes: A61M 2210/1458 Placenta
• CPC Class Codes: A61M 2210/1466 Umbilical cord
• 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 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 25/16 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 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 7/06 Antianaemics
• 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 2500/90 : Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2509/00 : Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 2533/90 : Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0068 : General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0605 : Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly

Abstract

The present invention provides a method of extracting and recovering embryonic-like stem cells, including, but not limited to pluripotent or multipotent stem cells, from an exsanguinated human placenta. A placenta is treated to remove residual umbilical cord blood by perfusing an exsanguinated placenta, preferably with an anticoagulant solution, to flush out residual cells. The residual cells and perfusion liquid from the exsanguinated placenta are collected, and the embryonic-like stem cells are separated from the residual cells and perfusion liquid. The invention also provides a method of utilizing the isolated and perfused placenta as a bioreactor in which to propagate endogenous cells, including, but not limited to, embryonic-like stem cells. The invention also provides methods for propagation of exogenous cells in a placental bioreactor and collecting the propagated exogenous cells and bioactive molecules therefrom.

Claims

1. An isolated mammalian placenta which has been exsanguinated and perfused under sterile conditions.

2. The isolated mammalian placenta of claim 1 wherein the solution used to perfuse the placenta contains an anticoagulant solution.

3. The isolated mammalian placenta of claim 1 wherein the solution used to perfuse the placenta contains an antimicrobial solution.

4. The isolated mammalian placenta of claim 1 wherein the solution used to perfuse the placenta contains growth factors.

5. The isolated mammalian placenta of claim 1 wherein the placenta is human.

6. The isolated mammalian placenta of claim 1 which has been stored from about 2 to 24 hours after the expulsion of the placenta from the uterus.

12. The isolated mammalian placenta of claim 1 which has been perfused for at least two hours.

13. The isolated mammalian placenta of claim 12 which has been perfused for at least eleven hours.

15. The isolated mammalian placenta of claim 13 which has been perfused for at least twenty-four hours.

7. An isolated mammalian placenta which has been exsanguinated and perfused and incubated under conditions to allow for the production of embryonic-like stem cells and other multipotent stem cells from said placenta.

8. The isolated mammalian placenta of claim 7 which has been incubated for a period of about 2 to 24 hours.

9. The isolated mammalian placenta of claim 7 which has been perfused or incubated for a period of about 24 to more than 48 hours.

17. The isolated mammalian placenta of claim 7 which has been perfused with a solution containing growth factors.

18. The isolated mammalian placenta of claim 7 which is human.

10. An isolated, perfused mammalian placenta which comprises viable embryonic-like stem cells.

11. The isolated placenta of claim 10 wherein the stem cells are OCT-4−

and ABC-p+.

14. The mammalian placenta of claim 10 wherein the placenta is recovered after birth.

16. The isolated mammalian placenta of claim 10 wherein the placenta is perfused with a solution containing growth factors.

19. A method of culturing a mammalian placenta comprising obtaining a placenta after expulsion from the uterus, exsanguinating the placenta, and perfusing the placenta under sterile conditions.

20. The method of claim 19 wherein the placenta is perfused with a solution containing an anticoagulant solution.

21. The method of claim 19 wherein the placenta is perfused with a solution containing an antimicrobial agent.

22. The method of claim 19 wherein the placenta is human.

23. The method of claim 19 wherein the placenta has been stored for about two to twenty-four hours after expulsion from the uterus.

24. The method of claim 19 wherein said expulsion is at birth.

25. The method of claim 23 wherein the placenta is stored at room temperature.

26. The method of claim 23 wherein the placenta is stored under refrigeration or freezer conditions.

27. A method for culturing an isolated mammalian placenta which has been exsanguinated and perfused comprising culturing the placenta under conditions to allow for the production of embryonic-like stem cells from said placenta.

28. The method of claim 27 wherein culturing the placenta comprises perfusing the placenta.

29. The method of claim 27 or 28 wherein the placenta has been incubated for a period of about twenty-four to forty-eight hours.

30. The method of claim 27 wherein the placenta has been perfused for at least two hours.

31. The method of claim 30 wherein the placenta has been perfused for at least eleven hours.

32. The method of claim 31 wherein the placenta has been perfused for at least twenty-four hours.

33. The method of claim 32 wherein the placenta has been preferred for more than forty-eight hours.

34. The method of claim 28 wherein the placenta has been perfused with a solution containing growth factors.

35. The method of claim 27 wherein the placenta is human.

37. The stem of claim 35 wherein the cell is a human cell.

39. The isolated placental stem cell of claim 35 wherein said cell is SSEA3−

and SEA4−

.

42. The apparatus of claim 39 further comprising a collection device for collecting stem cells.

43. The apparatus of claim 39 further comprising a means for monitoring and adjusting the culture conditions.

45. The apparatus of claim 39 further comprising a cell separation device.

46. A method of treating disease in a human which comprises administering a human placental stem of claim 35, 37 or 38.

47. A method of transplanting stem cells which comprises administering to a patient in need thereof a human placental stem cell of claim 35, 37 or 38.

48. A pharmaceutical composition which comprises a human placental stem cell of claim 35, 37 or 38.

49. The pharmaceutical composition of claim 46 which further comprises umbilical cord or placental blood.

50. A committed cell which has been differentiated from a human placental stem cell of claim 35, 37 or 38.

55. The stem cells of claim 49 or 51 wherein the stem cells originate from a placenta.

36. An isolated human placental stem cell which is OCT-4+ and ABC-p+.

38. An isolated mammalian placental stem cell having at least the following characteristics:

CD10+, CD29+, CD34−

, CD44+, CD45−

, CD54+, CD90+, SH2+, SH3+, SH4+, SSEA3−

, SSEA4−

, OCT-4+ and ABC-p+.

40. A human placental stem cell which has been isolated from a post-partum human placenta after said placenta has been exsanguinated and perfused for at least 11 hours.

41. A stem cell producing apparatus which comprises:

(a) a post-partum mammalian placenta which has been exsanguinated and perfused;

(b) a means for incubating or culturing the placenta; and

(c) a means for detecting stem cells.

44. The apparatus of claim 41 wherein the monitoring and adjusting is computerized.

51. An isolated, homogenous population of human placental stem cells which cap differentiate into all cell types.

52. A homogenous population of viable human placental stem cells which exhibit at least the following cell surface markers:

OCT-4+ and ABC-p+.

53. An isolated, homogenous population of human placental stem cells which is multipotent.

57. The composition of claim 53 further comprising cord blood having cells that are CD34+ and CD38+.

54. An isolated placenta containing a cell which is neither fetal nor maternal in origin.

59. The composition of claim 54 further comprising cord blood having cells that are CD34+ and CD38+.

56. A composition suitable for bone marrow transplantation which comprises a population of hematopoietic stem cells enriched in cells that are CD34+ and CD38−

.

58. A composition suitable for bone marrow transplantation which comprises a population of hematopoietic stem cells enriched in cells that are CD34−

and CD38−

.