Systems and methods for isolating and using clinically safe adipose derived regenerative cells

• US 8,163,276 B2
• Filed: 09/04/2009
• Issued: 04/24/2012
• Est. Priority Date: 12/07/2001
• Status: Expired due to Fees

• Current Assignee: Lorem Vascular Pte. Ltd.
• Original Assignee: Cytori Therapeutics Incorporated
• Inventors: Hedrick, Marc H., Fraser, John K., Schulzki, Michael J., Byrnes, Bobby, Carlson, Grace, Schreiber, Ronda, Wulur, Isabella
• Primary Examiner(s): Lankford, Jr., Blaine
• Application Number:
  US12/554,755
• Publication Number:
  US 20100136668A1
• Time in Patent Office: 963 Days
• Field of Search: None
• US Class Current: 424/93.7
• CPC Class Codes: A61B 17/00 Surgical instruments, devices or methods, e.g. tourniquets A61B18/00 takes precedence; contraceptive devices, pessaries, or applicators therefor A61F6/00; eye surgery A61F9/007; ear surgery A61F11/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 45/00 Means for pre-treatment of biological substances
• 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 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal 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 cell processing unit, comprising:

a tissue collection chamber that is configured to receive a first portion of unprocessed adipose tissue from a subject while maintaining a closed system, wherein said tissue collection chamber comprises an aspiration port which can be coupled to a suction device;

a first filter that is disposed within said tissue collection chamber, wherein said first filter is configured to substantially retain a first component of said unprocessed adipose tissue and substantially pass a second component of said unprocessed adipose tissue, such that said first filter substantially separates said first component from said second component, and wherein said first component comprises a cell population that comprises CD31 positive adipose-derived cells and said second component comprises lipid, blood, and mature adipocytes;

a processing chamber that is joined to said tissue collection chamber by a conduit such that a closed system is maintained, wherein said processing chamber comprises a cell concentrator and an outlet; and

a programmable processing device capable of communication with said tissue collection chamber and said processing chamber, wherein said processing device operates at least one pump configured to use pressure to control ingress and egress of tissues through said automated cell processing unit.

2. The automated cell processing unit of claim 1, wherein said cell concentrator is selected from the group consisting of a centrifuge, a filter, and a spinning membrane filter, or any combination thereof.

3. The automated cell processing unit of claim 2, further comprising:

a probe that can connect to at least one of said tissue collection chamber, said processing chamber, said cell concentrator, or a conduit that connects the tissue collection chamber and the cell processing chamber while maintaining a closed system.

4. The automated cell processing unit of claim 3, wherein said probe comprises a sensor.

5. The automated cell processing unit of claim 4, wherein said sensor is selected from the group consisting of an optical sensor, an ultrasonic sensor, and a pressure sensor.

6. The automated cell processing unit of claim 4, wherein said sensor is configured to detect the presence of red blood cells.

7. The automated cell processing unit of claim 2, wherein said probe is configured to remove a sample of said unprocessed adipose tissue, said first component of said first portion of unprocessed adipose tissue, said second component of said unprocessed adipose tissue, or a concentrated population of cells that comprise adipose-derived CD31-positive cells, while maintaining a closed system and transfer said sample to a testing chamber.

8. The automated cell processing unit of claim 7, wherein said testing chamber is configured to receive disaggregated adipose tissue, liberated CD31 positive adipose-derived cells, a biological fluid, or a concentrated population of cell comprising CD31 positive adipose-derived cells from the probe.

9. The automated cell processing unit of claim 7, wherein said testing chamber is configured to test said concentrated population of cells comprising adipose-derived CD31 positive cells for at least one factor selected from the group consisting of an endotoxin, a proteolytic enzyme, a platelet aggregating substance, and free lipids.

10. A method of processing adipose tissue to produce a concentrated population of cells comprising CD31 positive cells, comprising

introducing adipose tissue that comprises a population of cells comprising CD31positive cells into the automated cell processing unit of claim 1;

and

processing said adipose tissue to obtain a concentrated population of cells comprising adipose-derived CD31-positive cells.

11. The method of claim 10, wherein said automated cell processing until comprises a probe that comprises a sensor.

12. The method of claim 11, wherein said sensor is selected from the group consisting of an optical sensor, an ultrasonic sensor, and a pressure sensor.

13. The method of claim 11, wherein said sensor is configured to detect the presence of red blood cells in said sample.

14. The method of claim 10, further comprising:

receiving an input from a user that identifies whether said concentrated population of cells comprising adipose-derived CD31-positive cells will be administered intravascularly, non-systemically, or both; and

determining the presence of at least one factor selected from the group consisting adipocytes, free lipids, a proteolytic substance, a platelet aggregating substance, and an endotoxin in said concentrated population of cells comprising CD31-positive cells.

15. The method of claim 14, wherein the presence of an endotoxin, a proteolytic enzyme, a platelet aggregating substance, or free lipids is determined when the input received from said user identifies that said concentrated population of cells comprising CD31-positive cells will be administered intravascularly.

16. The method of claim 14, wherein said determining step is automatically performed.

17. The method of claim 14, wherein said determining step is not automatically performed.

18. The method of claim 10, further comprising:

receiving an input from a user that identifies the type of tissue desired to be formed from said concentrated population of cells comprising CD31-positive determining the presence of adipocytes, free lipids, a proteolytic substance, a platelet aggregating substance, and an endotoxin in said concentrated population of cells comprising CD31-positive cells.

19. The method of claim 18, wherein said determining step is automatically performed.

20. The method of claim 18, wherein said determining step is not automatically performed.

21. The method of claim 10, further comprising:

receiving an input from a user that identifies the type of tissue removed from said subject; and

determining the presence of adipocytes, free lipids, a proteolytic substance, a platelet aggregating substance, or an endotoxin in said concentrated population of cells comprising CD31-positive cells.

22. The method of claim 21, wherein said determining step is automatically performed.

23. The method of claim 21, wherein said determining step is not automatically performed.