Systems and methods for separating and concentrating regenerative cells from tissue

• US 20050084961A1
• Filed: 06/25/2004
• Published: 04/21/2005
• Est. Priority Date: 12/07/2001
• Status: Active Grant

• Current Assignee: Lorem Vascular Pte. Ltd.
• Original Assignee: Cytori Therapeutics Incorporated
• Inventors: Fraser, John K., Hedrick, Marc H., Schulzki, Michael J., Carlson, Grace, Byrnes, Bobby
• Granted Patent: US 7,514,075 B2
• Time in Patent Office: Days
• US Class Current: 435/366
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells

Abstract

Systems and methods are described that are used to separate cells from a wide variety of tissues. In particular, automated systems and methods are described that separate regenerative cells, e.g., stem and/or progenitor cells, from adipose tissue. The systems and methods described herein provide rapid and reliable methods of separating and concentrating regenerative cells suitable for re-infusion into a subject.

Claims

1. An automated system for separating and concentrating regenerative cells from tissue comprising:

a. a collection chamber to receive tissue removed from a patient, wherein the collection chamber facilitates disaggregation of the tissue such that regenerative cells are separated from the tissue; and

b. a processing chamber connected to the collection chamber to receive the disaggregated tissue from the collection chamber; and

c. a centrifugation device which can connect to the processing chamber, wherein the centrifugation device concentrates regenerative cells from the disaggregated tissue present in the processing chamber; and

d. a programmable processing device capable of communication with and controlling the collection chamber, the processing chamber, the centrifugation device and associated connections.

2. The system of claim 1, wherein the tissue is adipose tissue.

3. The system of claim 1, wherein the regenerative cells are stem cells.

4. The system of claim 1, wherein the regenerative cells are progenitor cells.

5. The system of claim 1, wherein the regenerative cells are a combination of stem cells and progenitor cells.

6. The system of claim 1, wherein the separated and concentrated regenerative cells are suitable for infusion into a patient.

7. The system of claim 6, wherein the regenerative cells are used to treat a patient.

8. The system of claim 1, wherein the collection chamber is further comprised of a filter housed in a filter cage within the collection chamber to retain tissue and filter out non-tissue components selected from the group consisting of free lipid, blood, and a washing solution.

9. The system of claim 1, wherein the collection chamber facilitates disaggregation of the tissue through agitation of a rotatable shaft comprised of paddles such that the paddles facilitate disaggregation of the tissue as the rotatable shaft is agitated.

10. The system of claim 9, wherein the paddles are attached to a filter cage within the collection chamber.

11. The system of claim 9, wherein the rotatable shaft is agitated through a magnetic stirrer.

12. The system of claim 1, wherein the collection chamber facilitates disaggregation of the tissue through rotation of a rotatable shaft comprised of paddles such that the paddles facilitate disaggregation of the tissue as the as the rotatable shaft is rotated.

13. The system of claim 9, wherein the rotatable shaft is rotated through a drive mechanism.

14. The system of claim 1, further comprising a washing solution source.

15. The system of claim 14, wherein the washing solution is saline.

16. The system of claim 1, further comprising a disaggregation agent source.

17. The system of claim 16, wherein the disaggregation agent is selected from the group consisting of collagenase, trypsin, lipase, hyaluronidase, deoxyribonuclease, Liberase H1, pepsin, or mixtures thereof.

18. The system of claim 1, wherein the processing chamber is comprised of one or more centrifuge chambers to hold the disaggregated tissue present in the processing chamber during centrifugation of the disaggregated tissue in the centrifuge device.

19. The system of claim 18, wherein the one or more centrifuge chambers further comprise one or more output chambers to collect the concentrated regenerative cells during centrifugation of the disaggregated tissue in the centrifuge device.

20. The system of claim 29, wherein the one or more output chambers are tilted at a different angle during centrifugation and a different angle for collection of the concentrated regenerative cells.

21. The system of claim 29, wherein the one or more output chamber can be manually re-positioned to arrive at the angle for collection of the concentrated regenerative cells.

22. The system of claim 21, wherein the concentrated regenerative cells are removed using a syringe.

23. The system of claim 1, wherein the processing chamber is comprised of a rotating seal to facilitate collection of the disaggregated tissue in a sterile manner.

24. The system of claim 23, wherein the rotating seal is comprised of a rotating shaft, two or more bearings, three or more lip seals, an outer housing, a circular channel and a circular spring.

25. The system of claim 23, wherein the rotating seal is comprised of a rotating shaft, a single rubber seal, an air gasket, one or more spring loaded seals, and two ceramic disks.

26. The system of claim 1, further comprising a waste chamber.

27. The system of claim 26, wherein the waste chamber receives the non-regenerative cell components of the tissue removed from the patient.

28. The system of claim 27, wherein the non-regenerative cell components present in the waste chamber are centrifuged to separate and concentrate components, including collagen, proteins, lipids, adipocytes and matrix components.

29. The system of claim 27, wherein the non-regenerative cell components present in the waste chamber are filtered to separate and concentrate components, including collagen, proteins, lipids, adipocytes and matrix components.

30. The system of claim 1, wherein the collection chamber and the processing chamber are connected through tubing.

31. The system of claim 30, wherein the tubing is further comprised of sensors.

32. The system of claim 1, wherein the collection chamber and the processing chamber further comprise one or more valves, pumps, sensors or combinations thereof.

33. The system of claim 30, wherein the collection chamber, the processing chamber and the tubing are disposable.

34. They system of claim 1, wherein the centrifuge device and the processing device are re-usable.

35. The system of claim 32, wherein the centrifugation device further comprises a centrifuge motor, a centrifuge motor controller and a centrifuge brake controller.

36. The processing device of claim 1, wherein the processing device controls the centrifuge chamber, the processing chamber and the centrifuge chamber by directing tissue flow through the use of one or more pumps.

37. The system of claim 36, wherein the pumps are peristaltic pumps.

38. The system of claim 37, wherein the pumps are re-usable.

39. The system of claim 1, wherein the processing device further comprises an user interface for an user to input parameters into the system.

40. The system of claim 1, wherein the processing device further comprises a display screen to display instructions that guide a user to input parameters, confirm pre-programmed steps, warn of errors or combinations thereof.

41. The system of claim 1, wherein a portion of the tissue removed from the patient is used to separate and concentrate regenerative cells and the remaining portion is set aside.

42. The system of claim 41, wherein the remaining portion of tissue is set aside in a sample chamber.

43. The system of claim 1, wherein one or more additives are added to the system.

44. The system of claim 43, wherein the additives are selected from growth factors, re-suspension fluids, cell culture reagents, cell expansion reagents, cell preservation reagents or cell modification reagents, or combinations thereof.

44-1. A system for culturing regenerative cells comprising:

a. removing tissue from a patient; and

b. providing a first system for separating and concentrating regenerative cells; and

c. providing a second system, wherein the second system is connected to the first system, for culturing the regenerative cells.

45. A system for genetically manipulating regenerative cells comprising:

a. removing tissue from a patient; and

b. providing a first system for separating and concentrating regenerative cells; and

c. providing a second system, wherein the second system is connected to the first system, for culturing the regenerative cells.

46. A system for separating and concentrating regenerative cells from tissue comprising:

a. a collection chamber to receive tissue removed from a patient, wherein the collection chamber facilitates disaggregation of the tissue such that regenerative cells are separated from the tissue; and

b. a processing chamber connected to the collection chamber to receive the disaggregated tissue from the collection chamber; and

c. at least one filter assembly, wherein the filter assembly concentrates the regenerative cells from the disaggregated tissue.

47. The system of claim 46, wherein the filter assembly comprises a percolative filtration device and a sedimentation device.

48. The system of claim 46, wherein the filter assembly comprises hollow fiber filtration device.

49. The system of claim 46, wherein the filter assembly comprises a percolative filtration device.

50. The system of claim 46, wherein the filter assembly comprises an elutriation device.