Isolation of spore-like cells from tissues exposed to extreme conditions

• US 20040175823A1
• Filed: 03/03/2004
• Published: 09/09/2004
• Est. Priority Date: 10/30/2000
• Status: Active Grant

• Current Assignee: VCellTherapeutics, Inc.
• Original Assignee: Vbi Technologies, LLC
• Inventors: Vacanti, Martin P., Vacanti, Charles A.
• Granted Patent: US 7,060,492 B2
• Time in Patent Office: Days
• US Class Current: 435/325
• 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: C12N 5/0602 Vertebrate cells

Abstract

Highly undifferentiated spore-like cells can be isolated from many different tissues and bodily fluids after those tissues and fluids have been exposed to extreme conditions. The spore-like cells can be used to treat a wide variety of disorders.

Claims

1. A method for isolating a spore-like cell from a biological tissue or fluid, the method comprising

(a) obtaining a tissue or fluid that has been exposed to an environment in which differentiated or partially differentiated cells in the tissue or fluid die and

(b) separating the spore-like cells from the differentiated or partially differentiated cells that have died.

2. The method of claim 1, further comprising disrupting the tissue or fluid either before or after step (a).

3. The method of claim 2, wherein disrupting the tissue or fluid comprises cutting the tissue into pieces, scraping the tissue with a blunt instrument, or passing the tissue or fluid through a series of devices having progressively smaller apertures.

4. The method of claim 3, wherein the devices are pipettes.

5. The method of claim 4, wherein the smallest pipette has an inner bore diameter of approximately 15 microns.

6. The method of claim 3, wherein the devices are filters.

7. The method of claim 6, wherein the finest filter has a pore size of approximately 15 microns.

8. The method of claim 1, wherein the biological tissue comprises a tissue that originates from the endoderm.

9. The method of claim 1, wherein the biological tissue comprises a tissue that originates from the mesoderm.

10. The method of claim 1, wherein the biological tissue comprises a tissue that originates from the ectoderm.

11. The method of claim 1, wherein the biological fluid comprises blood, urine, or saliva.

12. The method of claim 1, wherein the biological fluid is cerebrospinal fluid.

13. The method of claim 1, wherein the environment is an oxygen-poor environment.

14. The method of claim 1, wherein the environment is one in which the temperature is above or below the range of temperatures in which differentiated or partially differentiated cells can survive.

15. The method of claim 1, wherein the environment contains a toxin or infectious agent that kills differentiated or partially differentiated cells.

20. The method of claim 14, wherein the smallest pipette has an inner bore diameter of approximately 15 microns.

16. A method for isolating a spore-like cell from a biological tissue or fluid, the method comprising obtaining a tissue or fluid that has been exposed to an environment that is either 42°

C., or more, or 0°

C., or less, placing the tissue or fluid in a tissue culture vessel, allowing the spore-like cells to adhere to the vessel, and rinsing away non-spore-like cells, the tissue or fluid having been exposed to the environment without first being treated with a protective agent.

17. The method of claim 16, further comprising disrupting the tissue or fluid either before or after exposure to the environment or before or after placement in the tissue culture vessel.

18. The method of claim 17, wherein disrupting the tissue comprises cutting the tissue into pieces, scraping the tissue with a blunt instrument, or passing the tissue or fluid through a series of devices having progressively smaller apertures.

19. The method of claim 18, wherein the devices are pipettes.

21. The method of claim 18, wherein the devices are filters.

22. The method of claim 21, wherein the finest filter has a pore size of approximately 15 microns.

23. The method of claim 16, wherein the biological tissue or fluid originates from the endoderm.

24. The method of claim 16, wherein the biological tissue or fluid originates from the mesoderm.

25. The method of claim 16, wherein the biological tissue is a tissue that originates from the ectoderm.

26. The method of claim 16, wherein the biological fluid is blood, urine, or saliva.

27. The method of claim 16, wherein the biological fluid is cerebrospinal fluid.

28. The method of claim 16, wherein the environment is 0°

C. or less and the protective agent is a cryopreservative.

29. The method of claim 16, wherein the tissue or fluid is intentionally exposed to the environment.