Compositions and methods for generating skin

• US 20040033598A1
• Filed: 03/03/2003
• Published: 02/19/2004
• Est. Priority Date: 12/30/1999
• Status: Active Grant

• Current Assignee: VCellTherapeutics, Inc.
• Original Assignee: University Of Massachusetts
• Inventors: Vacanti, Martin P., Vacanti, Charles A.
• Granted Patent: US 7,560,275 B2
• Time in Patent Office: Days
• US Class Current: 435/371
• 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

Compositions and methods for growing new skin, which can be used to repair or replace skin that has been damaged or removed by a variety of insults.

Claims

1. An isolated spore-like cell, wherein the spore-like cell, when first isolated, is:

multipotent;

less than one to approximately seven microns in diameter;

and tolerant of oxygen-deprivation.

2. The spore-like cell of claim 1, wherein the cell has an outer coating containing sufficient glycolipids and mucopolysaccharides that the cell appears to have one or more dark stripes when viewed by transmission electron microscopy.

3. An isolated skin progenitor cell, wherein the skin progenitor cell is descended from a spore-like cell of claim 1.

4. The skin progenitor cell of claim 3, wherein the progenitor cell is isolated from the skin of a post-natal mammal.

5. The skin progenitor cell of claim 3, wherein the progenitor cell is isolated from a culture of spore-like cells.

6. The spore-like cell of claim 1, wherein the cell fails to express nestin.

7. The spore-like cell of claim 1, wherein the cell fails to express a component of keratin, tyrosinase, glutamic acid, glutamate, citrulline, tricohyaline, filagrin, epinephrine, norepinephrine, acetylcholine, or melanin granules.

8. The skin progenitor cell of claim 3, wherein the cell is isolated from the dermis of a post-natal mammal and, when placed in the vicinity of a skin wound in an animal, expresses a phenotype substantially similar to that of a sympathetic or parasympathetic neuron, or a cell within a sweat gland, a sebaceous gland, or a hair follicle.

9. The skin progenitor cell of claim 3, wherein the cell is isolated from the epidermis of a post-natal mammal and, when placed in the vicinity of a skin wound in an animal, expresses a phenotype substantially similar to that of a melanocyte, a keratinocyte, or a Merkel cell.

18. A skin construct comprising a cell of claim 1.

19. The skin construct of claim 18, further comprising a support structure.

20. The skin construct of claim 19, further comprising hydrogel, wherein the hydrogel and the cell of claim 1 form a hydrogel-spore-like cell composition.

21. A skin construct comprising a cell of claim 3.

22. The skin construct of claim 21, further comprising a support structure.

23. The skin construct of claim 22, further comprising hydrogel, wherein the hydrogel and the cell of claim 3 form a hydrogel-skin progenitor cell composition.

24. A method for generating artificial skin, the method comprising combining hydrogel with a spore-like cell of claim 1.

25. The method of claim 24, further comprising delivering the hydrogel-cell composition into a permeable, biocompatible support structure.

26. A method for generating artificial skin, the method comprising combining hydrogel with a skin progenitor cell of claim 3.

27. The method of claim 26, further comprising delivering the hydrogel-cell composition into a permeable, biocompatible support structure.

28. A method of treating damaged skin, the method comprising obtaining a hydrogel-spore-like cell composition of claim 20 and applying the composition to the damaged skin or to an area lying under the damaged skin.

29. A method of treating damaged skin, the method comprising obtaining a hydrogel-skin progenitor cell composition of claim 23 and applying the composition to the damaged skin or to an area lying under the damaged skin.

10. A method for isolating a spore-like cell or skin progenitor cell from a post-natal mammal, the method comprising passing a piece of skin obtained from the mammal through a series of devices having progressively smaller apertures.

11. The method of claim 10, wherein the device is a pipette.

12. The method of claim 11, wherein the smallest pipette in the series has an inner bore diameter of approximately 15 microns.

13. The method of claim 10, wherein the device is a filter.

14. The method of claim 10, wherein the finest filter in the series has a pore size of approximately 15 microns.

15. The method of claim 10 wherein the skin is exposed to a digestive enzyme.

16. The method of claim 15, wherein the digestive enzyme is trypsin or collagenase.

17. The method of claim 16, wherein the skin is incubated with approximately 0.05% trypsin at 37°

C. for approximately five minutes.

30. A spore-like cell or skin progenitor cell isolated by the method of claim 10.