Rapid preparation of stem cell matrices for use in tissue and organ treatment and repair

• US 7,906,110 B2
• Filed: 05/25/2005
• Issued: 03/15/2011
• Est. Priority Date: 02/23/2001
• Status: Active Grant

• Current Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Original Assignee: University of Pittsburgh of The Commonwealth System of Higher Education
• Inventors: Sacks, Michael S., Chancellor, Michael B., Huard, Johnny, Chung, Steve, Capelli, Christopher
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US11/138,168
• Publication Number:
  US 20050220775A1
• Time in Patent Office: 2,120 Days
• Field of Search: None
• US Class Current: 424/93.1
• 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: A61L 2430/22 for reconstruction of hollow organs, e.g. bladder, esophagus, urether, uterus
• CPC Class Codes: A61L 27/225 Fibrin; Fibrinogen
• CPC Class Codes: A61L 27/3629 Intestinal tissue, e.g. small intestinal submucosa
• CPC Class Codes: A61L 27/3679 Hollow organs, e.g. bladder, esophagus, urether, uterus, intestine
• CPC Class Codes: A61L 27/3834 Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 2533/56 Fibrin; Thrombin
• CPC Class Codes: C12N 2533/74 Alginate
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 2533/92 Amnion; Decellularised dermis or mucosa
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0658 Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources
• CPC Class Codes: C12N 5/0697 Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691

Abstract

The invention is directed to preparation of stem cell and physiologically acceptable matrix compositions. Compared with previous tissue engineering materials, the stem cell-matrix compositions of the present invention do not require long-term incubation or cultivation in vitro prior to use in vivo applications. The stem cells can be from numerous sources and may be homogeneous, heterogeneous, autologous, and/or allogeneic in the matrix material. The stem cell-matrix compositions provide point of service utility for the practitioner, wherein the stem cells and matrix can be combined not long before use, thereby alleviating costly and lengthy manufacturing procedures. In addition, the stem cells offer unique structural properties to the matrix composition which improves outcome and healing after use. Use of stem cells obtained from muscle affords contractility to the matrix composition.

Claims

1. A method of repairing or healing a wound in which the skin is damaged comprising:

a) isolating an end population of skeletal muscle-derived progenitor cells (MDCs) by a method comprising;

i) plating a suspension of cells from skeletal muscle tissue in a first container, to which fibroblast cells in the muscle tissue suspension adhere;

ii) re-plating non-adherent cells from i) in a second container after approximately 15% to 20% of the cells from the cell suspension have adhered to the first container;

iii) repeating step ii) at least one time to enrich for an end population of viable, non-fibroblast, desmin-expressing, skeletal muscle-derived myoblasts in the second container; and

iv) isolating an end population of viable, non-fibroblast, desmin-expressing skeletal muscle-derived myoblasts in the culture;

b) admixing a preparation comprising the MDCs with a physiologically acceptable matrix material to form a stem cell matrix dressing;

c) incubating the stem cell matrix dressing in vitro for less than about 12 hours prior to use in dressing a wound in which the skin is damaged; and

d) dressing said wound in which the skin is damaged with the stem cell matrix, thereby repairing or healing said wound.

2. The method according to claim 1, wherein the MDCs are autologous to a recipient.

3. The method according to claim 1, wherein the MDCs are allogeneic to a recipient.

4. The method according to claim 1, wherein the physiologically acceptable matrix material is absorbable.

5. The method according to claim 1, wherein the physiologically acceptable matrix material is non-absorbable.

6. The method according to claim 1, wherein the physiologically acceptable matrix material is selected from the group consisting of small intestine submucosa (SIS), crosslinked alginate, bioadhesives, hydrocolloid, collagen gel, collagen sponge, polyglycolic acid (PGA) mesh, polyglactin (PGL) mesh, fleeces, and dead de-epidermized skin equivalents.

7. The method according to claim 1, wherein the stem cell matrix dressing is contractible.

8. The method according to claim 1, wherein about 2.5×

10³ to about 1×

10⁶ MDCs are admixed with the physiologically acceptable matrix material.

9. The method according to claim 1, wherein about 5×

10³ to about 1×

10⁶ MDCs are admixed with the physiologically acceptable matrix material.

10. The method according to claim 1, wherein about 1×

10⁵ MDCs are admixed per 1 cm² of the physiologically acceptable matrix material.

11. The method according to claim 1, wherein the stem cell matrix dressing is incubated in vitro for less than about 3 hours prior to use in dressing said wound in which the skin is damaged.

12. The method according to claim 1, wherein the stem cell matrix dressing is incubated in vitro for less than about 1 hour prior to use in dressing said wound in which the skin is damaged.

13. The method according to claim 1, wherein the stem cell matrix dressing is incubated in vitro for less than about 30 minutes prior to use in dressing said wound in which the skin is damaged.

14. The method according to claim 1, wherein the MDCs stem cell matrix dressing is incubated in vitro from about 5 seconds to about 30 minutes prior to use in dressing said wound in which the skin is damaged.

15. The method according to claim 1, wherein dressing said wound comprises applying the stem cell matrix to the skin tissue by spraying, painting, coating, or spreading.

16. A method of repairing or healing a wound in which the skin is damaged comprising:

a) admixing a preparation comprising desmin expressing skeletal muscle-derived progenitor cells with a physiologically acceptable matrix material to form a stem cell matrix dressing; and

b) incubating the stem cell matrix dressing in vitro for less than about 12 hours prior to use in dressing a wound in which the skin is damaged; and

c) dressing said wound in which the skin is damaged with the stem cell matrix, thereby repairing or healing said wound.

17. The method according to claim 16, wherein the desmin expressing skeletal muscle-derived progenitor cells are autologous to a recipient.

18. The method according to claim 16, wherein the desmin expressing skeletal muscle-derived progenitor cells are allogeneic to a recipient.

19. The method according to claim 16, wherein the physiologically acceptable matrix material is absorbable.

20. The method according to claim 16, wherein the physiologically acceptable matrix material is non-absorbable.

21. The method according to claim 16, wherein the physiologically acceptable matrix material is selected from the group consisting of small intestine submucosa (SIS), crosslinked alginate, bioadhesives, hydrocolloid, collagen gel, collagen sponge, polyglycolic acid (PGA) mesh, polyglactin (PGL) mesh, fleeces, and dead de-epidermized skin equivalents.

22. The method according to claim 16, wherein the stem cell matrix dressing is contractible.

23. The method according to claim 16, wherein about 2.5×

10³ to about 1×

10⁶ desmin expressing skeletal muscle-derived progenitor cells are admixed with the physiologically acceptable matrix material.

24. The method according to claim 16, wherein about 5×

10³ to about 1×

10⁶ desmin expressing skeletal muscle-derived progenitor cells are admixed with the physiologically acceptable matrix material.

25. The method according to claim 16, wherein about 1×

10⁵ desmin expressing skeletal muscle-derived progenitor cells are admixed per 1 cm² of the physiologically acceptable matrix material.

26. The method according to claim 16, wherein the stem cell matrix dressing is incubated in vitro for less than about 3 hours prior to use in dressing said wound in which the skin is damaged.

27. The method according to claim 16, wherein the stem cell matrix dressing is incubated in vitro for less than about 1 hour prior to use in dressing said wound in which the skin is damaged.

28. The method according to claim 16, wherein the stem cell matrix dressing is incubated in vitro for less than about 30 minutes prior to use in dressing said wound in which the skin is damaged.

29. The method according to claim 16, wherein the stem cell matrix dressing is incubated in vitro from about 5 seconds to about 30 minutes prior to use in dressing said wound in which the skin is damaged.

30. The method according to claim 16, wherein dressing said wound comprises applying the stem cell matrix to the skin tissue by spraying, painting, coating, or spreading.