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

• US 7,585,670 B2
• Filed: 07/02/2004
• Issued: 09/08/2009
• 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., Wulur, Isabella, Schulzki, Michael J., Carlson, Grace, Schreiber, Ronda Elizabeth, Byrnes, Bobby
• Primary Examiner(s): LANKFORD JR, LEON B
• Application Number:
  US10/884,638
• Publication Number:
  US 20050260175A1
• Time in Patent Office: 1,894 Days
• Field of Search: None
• US Class Current: 435/325
• 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. A method of processing adipose tissue that comprises a population of cells comprising adipose-derived stem cells for reintroduction into a subject, comprising:

introducing adipose tissue that comprises said population of cells comprising adipose-derived stem cells removed from said subject into a partially or fully automated self-contained adipose-derived stem cell processing unit, wherein said automated cell processing unit comprises;

a tissue collection chamber that is configured to receive unprocessed adipose tissue that is removed from said patient, wherein said tissue collection chamber is defined by a closed system;

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

a processing chamber which is configured to receive said first component comprising a cell population that comprises adipose-derived stem cells from said tissue collection chamber, wherein said processing chamber is within said closed system;

a conduit configured to allow passage of said first component comprising a cell population comprising adipose-derived stem cells from said tissue collection chamber to said processing chamber while maintaining a closed system;

a cell concentrator disposed within said processing chamber, which is configured to facilitate the concentration of said first component comprising a cell population that comprises adipose-derived stem cells so as to obtain a concentrated population of cells that comprise adipose-derived stem cells, wherein said cell concentrator comprises a centrifuge or a spinning membrane filter;

an outlet configured to allow the aseptic removal of said concentrated population of cells that comprise adipose-derived stem cells;

a probe that can connect to at least one of said tissue collection chamber, said processing chamber, said conduit or said concentrating device, wherein said probe is configured to remove a sample of said unprocessed adipose tissue, said first component of said unprocessed adipose tissue, said second component of said unprocessed adipose tissue, or said concentrated population of cells that comprise adipose-derived stem cells, while maintaining a closed system and transfer said sample to a testing chamber;

said testing chamber that can receive said sample from said probe; and

a programmable processing device capable of communicating with and controlling at least one part of the self-contained adipose-derived stem cell processing unit selected from the group consisting of the tissue collection chamber, the processing chamber, the cell concentrator, the probe, and the testing chamber;

processing said adipose tissue to obtain a concentrated population of cells comprising adipose-derived stem cells; and

performing at least one test for at least one factor on said sample.

2. The method of claim 1, wherein said probe comprises a sensor.

3. The method of claim 2, wherein said sensor is selected from the group consisting of an optical sensor, an ultrasonic sensor, and a pressure sensor.

4. The method of claim 2, wherein said sensor is configured to detect the presence of red blood cells in said sample.

5. The method of claim 1, wherein said testing further comprises:

receiving an input identifying whether said concentrated population of cells that comprises adipose-derived stem cells will be reintroduced into said subject intravascular, non-systemically, or both; and

selecting said at least one test for at least one factor selected from the group consisting adipocytes, free lipids, a proteolytic substance, a platelet aggregating substance, and an endotoxin based upon said input.

6. The method of claim 5, wherein said testing comprises testing said concentrated population of cells comprising adipose-derived stem cells for at least one factor selected from the group consisting of an endotoxin, a proteolytic enzyme, a platelet aggregating substance, and free lipids, when said input comprises intravascular reintroduction.

7. The method of claim 5, wherein said selecting step is automatically performed by said automated self contained adipose-derived stem cell processing unit.

8. The method of claim 5, wherein said selecting step is not automatically performed by said automated self contained adipose-derived stem cell processing unit.

9. The method of claim 1, wherein said testing further comprises:

receiving an input identifying the type of tissue desired to be formed from said concentrated population of cells comprising adipose-derived stem cells; and

selecting said at least one test for at least one factor based upon said input, wherein said at least one factor is selected from the group consisting of adipocytes, free lipids, a proteolytic substance, a platelet aggregating substance, and an endotoxin.

10. The method of claim 9, wherein said selecting step is automatically performed by said automated self contained adipose-derived stem cell processing unit.

11. The method of claim 9, wherein said selecting step is not automatically performed by said automated self contained adipose-derived stem cell processing unit.

12. The method of claim 1, wherein said testing further comprises:

receiving an input identifying the type of tissue removed from said subject; and

selecting said at least one test for at least one factor based upon said input, wherein said at least one factor is selected from the group consisting of adipocytes, free lipids, a proteolytic substance, a platelet aggregating substance, and an endotoxin.

13. The method of claim 12, wherein said selecting step is automatically performed by said automated self contained adipose-derived stem cell processing unit.

14. The method of claim 12, wherein said selecting step is not automatically performed by said automated self contained adipose-derived stem cell processing unit.

15. The method of claim 1, wherein said cell concentrator comprises a centrifuge.

16. The method of claim 1, wherein said cell concentrator comprises a spinning membrane filter.

17. The method of claim 1, wherein said processing step involves enzymatic disaggregation of said tissue.

18. The method of claim 1, wherein the tissue comprises adipose tissue.

19. The method of claim 1, wherein said concentrated population of cells comprising adipose-derived stem cells comprises progenitor cells.

20. The method of claim 1, wherein said cell concentrator comprises a filter.

21. The method of claim 1, wherein said self contained adipose-derived stem cell processing unit farther comprises a testing chamber that can receive said sample of disaggregated adipose tissue, liberated regenerative cells, biological fluid, or concentrated cell population of cells comprising adipose-derived stem cells from said probe.