SYSTEMS, METHODS AND COMPOSITIONS FOR OPTIMIZING TISSUE AND CELL ENRICHED GRAFTS

• US 20100279405A1
• Filed: 04/30/2010
• Published: 11/04/2010
• Est. Priority Date: 05/01/2009
• Status: Active Grant

• Current Assignee: Lorem Vascular Pte. Ltd.
• Original Assignee: Bimini Technologies LLC
• Inventors: Peterson, Alvin, Fornace, Lucas
• Granted Patent: US 9,133,431 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 35/35 Fat tissue; Adipocytes; Stromal cells; Connective tissues adipose-derived stem cells A61K35/28; collagen A61K38/39
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12M 45/03 by control of the humidity or content of liquids; Drying
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue

Abstract

Disclosed herein are methods and systems for the concentration of cells from a cell suspension into unprocessed tissue, such as adipose tissue. Also disclosed herein are systems for optimizing hydration of tissue and cell enriched grafts.

Claims

1. A device for preparing tissue for an adipose tissue graft, comprising:

a flexible, collapsible bag having a first chamber and a second chamber, which are defined by a filter comprising pores;

a separator located within the second chamber;

an inlet port connected to the flexible, collapsible bag, wherein said inlet port is configured to allow the aseptic introduction of adipose tissue into the first chamber; and

an outlet port connected to the flexible, collapsible bag, wherein the outlet port is configured to aseptically remove liquid and cells from the second chamber.

2. The device of claim 1, wherein the separator is a free floating porous structure within said second chamber.

3. The device of claim 1, wherein the separator is a porous structure that defines a third chamber within the second chamber.

4. The device of claim 1, wherein the bag comprises a first outer shell and a second outer shell joined by a double heat seal.

5. The device of claim 4, wherein the filter is joined to the first outer shell and the second outer shell by a double heat seal.

6. The device of claim 1, wherein the separator comprises a lipid-wicking material.

7. The device of claim 6, wherein said lipid-wicking material is a polyester mesh screen.

8. The device of claim 1, wherein said separator comprises a plurality of pores that are larger than the pores of said filter.

9. The device of claim 8, wherein the pore size of said second filter is greater than or equal to about 5, 6, 7, 8, 9, or 10 times larger than the pore size of said filter.

10. The device of claim 1, wherein the pore size of said filter is greater than or equal to about 30 μ

M.

11. The device of claim 1, wherein the pore size of said filter is between about 30 μ

M and about 200 μ

M.

12. The device of claim 1, wherein the separator comprises pores and wherein the pore size of said separator is between about 300 and 2000 μ

M.

13. The device of claim 1, wherein said inlet port is configured to releasably connect with an adapter, said adapter being configured to releasably connect with the tip of a syringe barrel.

14. The device of claim 13, wherein the inlet port is configured to allow material to enter into the port, but not to exit from the port.

15. The device of claim 14, wherein said inlet port comprises a deformable plastic valve.

16. The device of claim 13, wherein the syringe barrel is a 60 mL catheter syringe barrel.

17. The device of claim 1, wherein the inlet port is configured to be attached to a cannula, while maintaining a sterile fluid/tissue pathway.

18. The device of claim 1, further comprising a second device, said second device comprising an adipose-derived regenerative cell isolation device attached to said device.

19. The device of claim 18, wherein said second device comprises a device of claim 1.

20. The device of claim 18, wherein the second device is connected to the device of claim 1 by a conduit that is configured to transfer isolated adipose-derived regenerative cells from said second device to the first chamber of the device of claim 1.

21. The device of claim 20, wherein said conduit comprises a Y connection.

22. A method of making an adipose tissue graft, comprising:

(a) obtaining a first portion of unprocessed adipose tissue;

(b) introducing said first portion of unprocessed adipose tissue into the first chamber of a first device of claim 1;

(c) rinsing the first portion of unprocessed adipose tissue by adding a physiologic wash solution to the first chamber; and

(d) removing fluid from the second chamber of the first device, wherein the fluid is selected from the group consisting of water, physiologic wash solution, blood and free lipid, or any combination thereof, thereby drying the adipose tissue.

23. The method of claim 22, wherein the physiologic solution is selected from the group consisting of Lactated Ringer'"'"'s solution, Hartmann'"'"'s Solution, saline, and PLASMALYTE™

solution.

24. The method of claim 22, wherein said dried adipose tissue of (c) is dehydrated to a liquid content that is less than about ½

, ⅓

, or ¼

times that of said first portion unprocessed adipose tissue prior to the introducing step.

25. The method of claim 22, wherein said dried adipose tissue of (c) is dehydrated to a liquid content that is less than about ⅓

that of said first portion unprocessed adipose tissue prior to the introducing step.

26. The method of claim 22, further comprising isolating a population of adipose-derived regenerative cells from a second portion of adipose tissue and contacting the dehydrated adipose tissue of (d) with the isolated population of adipose-derived regenerative cells under conditions that allow the isolated population of adipose-derived regenerative cells to permeate the dried adipose tissue of (d).

27. The method of claim 26, wherein said isolated population of adipose-derived regenerative cells is not subjected to centrifugation prior to contacting said dehydrated adipose tissue of (c).

28. The method of claim 26, wherein said isolated population of adipose-derived regenerative cells is prepared in a second device of claim 1 by contacting adipose tissue present in said first chamber of said second device with collagenase under conditions that liberate said cells.

29. The method of claim 28, wherein contacting the dehydrated adipose tissue of (d) with said isolated population of adipose-derived regenerative cells is performed in the first device.