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

• US 9,598,669 B2
• Filed: 12/28/2006
• Issued: 03/21/2017
• Est. Priority Date: 12/29/2005
• Status: Active Grant

• Current Assignee: Celularity, Inc.
• Original Assignee: Anthrogenesis Corporation (Bristol-Myers Squibb Company)
• Inventors: Edinger, James W., Heidaran, Mohammad, Hofgartner, Wolfgang
• Primary Examiner(s): HANLEY, SUSAN MARIE
• Application Number:
  US11/648,812
• Publication Number:
  US 20070190042A1
• Time in Patent Office: 3,736 Days
• US Class Current: 1/1
• 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 composition comprising, in a physiologically-acceptable solution, a JNK inhibitor, wherein said JNK inhibitor is 3-(4-fluoro-phenyl)-1H-indazole-5-carboxylic acid amide;

a protease;

an oxygen-carrying perfluorocarbon;

3-(4-amino-1-oxo-1,3-dihydroisoindol-2-yl)-piperidine-2,6-dione;

atrial natriuretic peptide (ANP); and

either pyrrolidine dithiocarbamate or clonazepam;

wherein said composition is not a naturally-occurring composition, and wherein said JNK inhibitor is an inhibitor of apoptosis; and

wherein the composition is suitable for collecting placental stem cells.

2. The composition of claim 1, wherein said protease is present in an amount sufficient to detectably dissociate cells of a tissue comprising stem cells.

3. The composition of claim 1 wherein said physiologically-acceptable solution is a saline solution or culture medium.

4. The composition of claim 3 wherein said saline solution is 0.9% NaCl solution or phosphate buffered saline.

5. The composition of claim 1, wherein said protease is a matrix metalloprotease or a neutral protease.

6. The composition of claim 5, wherein said matrix metalloprotease is collagenase.

7. The composition of claim 5, wherein said neutral protease is thermolysin or dispase.

8. The composition of claim 1, wherein said composition comprises hyaluronidase.

9. The composition of claim 1, additionally comprising hydroxyethyl starch, lactobionic acid and raffinose.

10. The composition of claim 1, additionally comprising UW solution.

11. The composition of claim 1, wherein said JNK inhibitor is an indazole.

12. The composition of claim 1 comprising a plurality of stem cells.

13. The composition of claim 12, wherein said stem cells comprise CD34⁺ stem cells.

14. The composition of claim 13, wherein said CD34⁺ stem cells comprise CD34⁺CD38⁻

stem cells.

15. The composition of claim 13, wherein said CD34⁺CD38⁻

stem cells are part of a population of CD34⁺CD38⁻

stem cells present in placental perfusate as a higher percentage of total nucleated cells as compared to the percentage of CD34⁺CD38⁻

cells in cord blood.

16. The composition of claim 12, wherein said stem cells are CD34⁻

stem cells.

17. The composition of claim 16, wherein said CD34⁻

stem cells are additionally HLA-G⁺ or CD200⁺.

18. The composition of claim 16, wherein said CD34⁻

stem cells are additionally OCT-4⁺, CD73⁺, or CD105+.