Method for processing lipoaspirate cells
View Patent ↗Cells present in processed lipoaspirate tissue are used to treat patients. Methods of treating patients include processing adipose tissue to deliver a concentrated amount of stem cells obtained from the adipose tissue to a patient. The methods may be practiced in a closed system so that the stem cells are not exposed to an external environment prior to being administered to a patient. Compositions that are administered to a patient include a mixture of adipose tissue and stem cells so that the composition has a higher concentration of stem cells than when the adipose tissue was removed from the patient.
1. A method of processing a cell population that comprises adipose-derived stem cells, comprising:
removing a first portion of adipose tissue that comprises a cell population that comprises adipose-derived stem cells from a patient;
introducing the removed first portion of adipose tissue that comprises said cell population that comprises adipose-derived stem cells into a self-contained adipose-derived stem cell processing unit configured to maintain a closed pathway, wherein said self-contained adipose derived stem cell processing unit comprises:
a tissue collection container that is configured to receive adipose tissue that is removed from a patient, wherein said tissue collection container is defined by a closed system;
a first filter that is disposed within said tissue collection container, 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, mature adipocytes, and saline;
a cell collection container, which is configured to receive said first component comprising a cell population that comprises adipose-derived stem cells from said tissue collection container, wherein said cell collection container 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 container to said cell collection container while maintaining a closed system;
a cell concentrator disposed within said cell collection container, 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 comprises adipose-derived stem cells, wherein said cell concentrator comprises a centriftige or a spinning membrane filter; and
an outlet configured to allow the aseptic removal of said concentrated population of cells that comprise adipose-derived stem cells;
separating and concentrating said cell population that comprises adipose-derived stem cells from said removed first portion of adipose tissue within said self-contained cell processing unit while maintaining said closed pathway to obtain a concentrated cell population that comprises adipose-derived stem cells; and
mixing said concentrated cell population that comprises adipose-derived stem cells with a second portion of unprocessed adipose tissue from said patient.
2. The method of claim 1 ,wherein said first or second portion of adipose tissue that is removed from said patient is lipoaspirate.
3. The method of claim 1 , wherein said first or second portion of adipose tissue that is removed from said patient is obtained by excisional lipectomy.
4. The method of claim 1 , further comprising a disaggregation step, wherein said cell population that comprises adipose-derived stem cells contained in said removed first portion of adipose tissue is mechanically or enzymatically disaggregated from said mature adipocytes and connective tissue present in said first portion of adipose tissue that was removed from said patient prior to separation.
5. The method of claim 1 , wherein said cell concentrator comprises a spinning membrane filter.
6. The method of claim 1 , wherein said cell concentrator comprises an antibody.
7. The method of claim 6 , wherein said antibody is selected from the group consisting of AP2, CD3, CD 19, and CD11 b.
8. The method of claim 1 , wherein said cell concentrator comprises a centrifuge.
9. The method of claim 4 , wherein said disaggregation step comprises an enzymatic digestion.
10. The method of claim 9 , wherein said enzymatic digestion comprises a collagenase.
11. The method of claim 9 , wherein said enzymatic digestion comprises a neutral protease.
12. The method of claim 9 , wherein said enzymatic digestion comprises trypsin.
13. The method of claim 1 , wherein said adipose-derived stem cells in said concentrated cell population that comprises adipose-derived stem cells are at least 0.1% of the cellular component.
14. The method of claim 1 , wherein said adipose-derived stem cells in said concentrated cell population that comprises adipose-derived stem cells are between about 2% and about 12% of the cellular component.
15. The method of claim 1 , wherein said concentrated cell population that comprises adipose-derived stem cells has a concentration of about 2 ×10 7 cells/100 ml of adipose tissue.
16. The method of claim 1 , wherein said concentrated cell population that comprises adipose-derived stem cells comprises endothelial precursor cells.
17. The method of claim 1 , further comprising contacting said concentrated cell population that comprises adipose-derived stem cells with an additive.
18. The method of claim 17 , wherein said additive is a tissue or tissue fragment.
19. The method of claim 18 , wherein said tissue or tissue fragment is unprocessed adipose tissue.
20. The method of claim 18 , wherein said tissue or tissue fragment is a connective tissue or bone.
21. The method of claim 17 , wherein said additive is a biological or artificial scaffold.