Composition for collecting and preserving placental stem cells and methods of using the composition

• US 9,725,694 B2
• Filed: 07/29/2010
• Issued: 08/08/2017
• Est. Priority Date: 12/29/2005
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Heidaran, Mohammad, Edinger, James W., Hofgartner, Wolfgang
• Primary Examiner(s): Hanley, Susan
• Application Number:
  US12/846,765
• Publication Number:
  US 20100291679A1
• Time in Patent Office: 2,567 Days
• Field of Search: None
• 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. A method of isolating stem cells, the method comprising contacting said stem cells with a solution comprising a caspase inhibitor, a JNK inhibitor, a protease, and either pyrrolidine dithiocarbamate or clonazepam,

wherein the JNK inhibitor is an inhibitor of apoptosis, said JNK inhibitor represented by the structure

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

3. The method of claim 1, additionally comprising contacting said stem cells with an oxygen-carrying perfluorocarbon.

4. The method of claim 1, wherein said solution is a saline solution.

5. The method of claim 1, wherein said solution additionally comprises hydroxyethyl starch, lactobionic anion and raffinose.

6. The method of claim 1, wherein said solution comprises UW solution.

7. The method of claim 1, wherein said perfusion is performed by passing said perfusion solution into one or both of the umbilical artery and umbilical vein of said placenta.

8. The method of claim 1, wherein said perfusion solution comprises 0.9% NaCl solution or phosphate buffered saline.

9. The method of claim 1, wherein said perfusion uses from about 100 mL to about 10,000 mL of said perfusion solution.

10. The method of claim 1, said method comprising isolating said stem cells, 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.

11. The method of claim 1, additionally comprising contacting the stem cells with an amino-substituted isoindoline.

12. The method of claim 11, wherein said amino-substituted isoindoline is a compound represented by the structure

13. The method of claim 11, wherein said substituted isoindoline 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.

14. The method of claim 1, wherein said solution additionally comprises a vasodilator.

15. The method of claim 14, wherein said vasodilator is atrial natriuretic peptide (ANP), adrenocorticotropin, corticotropin-releasing hormone, sodium nitroprusside, hydralazine, adenosine triphosphate, adenosine, indomethacin or magnesium sulfate.

16. The method of claim 1, wherein said stem cells are CD34+ stem cells.

17. The method of claim 1, wherein said stem cells are tissue culture plastic-adherent CD34⁻

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

18. The method of claim 1, wherein said solution is culture medium.