COMPOSITION FOR COLLECTING AND PRESERVING PLACENTAL STEM CELLS AND METHODS OF USING THE COMPOSITION

• US 20100291679A1
• Filed: 07/29/2010
• Published: 11/18/2010
• Est. Priority Date: 12/29/2005
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Hofgartner, Wolfgang, Edinger, James W., Heidaran, Mohammad
• Granted Patent: US 9,725,694 B2
• Time in Patent Office: Days
• US Class Current: 435/374
• CPC Class Codes: C12N 2501/48 Regulators of apoptosis
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• 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 improved compositions and methods for the collection of stem cells from an organ, e.g., placenta. The invention provides a stem cell collection composition comprising an apoptosis inhibitor and, optionally, an enzyme such as a protease or mucolytic enzyme, vasodilator, necrosis inhibitor, oxygen-carrying perfluorocarbon, or an organ preserving compound. The invention provides methods of using the stem cell collection composition to collect stem cells and to preserve populations of stem cells.

Claims

1-35. -35. (canceled)

36. A method of isolating stem cells or preserving a population of stem cells, comprising contacting said stem cells with a solution comprising a caspase inhibitor, a JNK inhibitor and a protease, and isolating said stem cells,

wherein JNK inhibitor is an inhibitor of apoptosis, said JNK inhibitor having the structure

37. The method of claim 36, wherein the JNK inhibitor is 3-(4-fluoro-phenyl)-1H-indazole-5-carboxylic acid amide.

38. The method of claim 36, additionally comprising contacting said stem cells with a compound that inhibits necrosis.

39. The method of claim 36, additionally comprising contacting said stem cells with an oxygen-carrying perfluorocarbon.

40. The method of claim 36, wherein said solution is a saline solution or culture medium.

41. The method of claim 36, wherein said solution additionally comprises hydroxyethyl starch, lactobionic anion and raffinose.

42. The method of claim 36, wherein said solution comprises UW solution.

43. The method of claim 36, wherein said stem cells are isolated from mammalian placenta, umbilical cord, placental blood or umbilical cord blood.

44. The method of claim 36, wherein said stem cells are isolated from a tissue by physical or enzymatic disruption of said tissue.

45. The method of claim 43, wherein said stem cells are isolated from said mammalian placenta, and said isolating is performed by perfusing said mammalian placenta with a perfusion solution comprising said caspase inhibitor, said JNK inhibitor, and said protease.

46. The method of claim 45, wherein said perfusing is performed by passing said perfusion solution into one or both of the umbilical artery and umbilical vein of said placenta.

47. The method of claim 45, wherein said perfusion solution comprises 0.9% NaCl solution or phosphate buffered saline.

48. The method of claim 45, wherein said perfusing uses from about 100 mL to about 10,000 mL of said perfusion solution.

49. The method of claim 36, wherein said stem cells are exposed to a hypoxic condition for less than 24 hours during said isolation, wherein a hypoxic condition is a concentration of oxygen that is less than normal blood oxygen concentration.

50. The method of claim 36, additionally comprising contacting the stem cell with amino-substituted isoindoline.

51. The method of claim 36, wherein said substituted isoindoline is a compound having the structure

52. The method of claim 36, wherein said substituted thalidomide is 3-(4-amino-1-oxo-1,3-dihydroisoindol-2-yl)-piperidine-2,6-dione, or 4-(Amino)-2-(2,6-dioxo(3-piperidyl))-isoindoline-1,3-dione.

53. The method of claim 36, wherein said solution additionally comprises a vasodilator.

54. The method of claim 53, wherein said vasodilator is atrial natriuretic peptide (ANP), adrenocorticotropin, corticotropin-releasing hormone, sodium nitroprusside, hydralazine, adenosine triphosphate, adenosine, indomethacin or magnesium sulfate.

55. The method of claim 36, wherein said stem cells are CD34⁺ stem cells.

56. The method of claim 36, wherein said stem cells are tissue culture plastic-adherent CD34⁻

, CD10⁺, CD105⁺, CD200⁺ placental stem cells.