Methods of mediating macrophage phenotypes

Patent metadata

Field	Value
Publication number	US-9833474-B2
Application number	US-201414554381-A
Country	US
Kind code	B2
Filing date	Nov 26, 2014
Priority date	Nov 26, 2013
Publication date	Dec 5, 2017
Grant date	Dec 5, 2017

Abstract

Methods of inducing a polarization of macrophages. The method includes obtaining a blood fraction, fractionating the blood fraction to produce a blood fraction, and contacting the blood fraction with a source of macrophages. A blood fraction including platelet-poor plasma polarizes the source of macrophages into M1 macrophages. A blood faction including a protein solution polarizes the source of macrophages into M2 macrophages.

First claim

What is claimed is:

1. A method for mediating the phenotype of macrophages, consisting of:
(a) obtaining a blood-derived composition; and
(b) contacting an isolated source of macrophages with the blood-derived composition and optionally a non-differentiating growth media, wherein contacting the macrophages with the blood-derived composition induces the macrophages to a polarized phenotype.

2. The method according to claim 1 , wherein the source of macrophages is a composition comprising monocytes.

3. The method according to claim 1 , wherein the blood-derived composition is platelet-poor plasma, and wherein the source of macrophages is polarized to M1 macrophages.

4. The method according to claim 1 , wherein the blood-derived composition is a protein solution comprising interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-1 receptor antagonist (IL-1ra), and transforming growth factor-β (TGF-β), and wherein the source of macrophages are polarized to M2 macrophages.

5. The method according to claim 1 , wherein step (a) consists of contacting whole blood, bone marrow aspirate, a fraction thereof, a concentrate thereof, or a combination thereof, with polyacrylamide beads to form an activating composition, and separating the activating composition from the polyacrylamide beads to produce the blood-derived composition.

6. The method according to claim 1 , wherein the blood-derived composition comprises whole blood, platelet rich plasma, bone marrow aspirate, a bone marrow concentrate, a protein solution, or platelet poor plasma, a fraction thereof, a concentrate thereof, or a combination thereof.

7. The method according to claim 1 , wherein the source of macrophages is peripheral blood, tissue at or near a site of inflammation in a donor, or a culture medium comprising monocytes.

8. The method according to claim 1 , wherein the source of macrophages is a concentrated macrophage solution generated by fractionating peripheral blood or bone marrow aspirate obtained from a donor.

9. A method for inducing M2 polarization of macrophages, consisting of:
(a) obtaining a blood-derived composition comprising interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-1 receptor antagonist (IL-1ra), transforming growth factor β-1 (TGFβ-1), or a combination thereof; and
(b) contacting an isolated source of macrophages with the blood-derived composition, wherein contacting the macrophages with the blood-derived composition induces the macrophages to polarize to an M2 phenotype.

10. The method according to claim 9 , wherein the blood-derived composition consists of whole blood, platelet rich plasma, a protein solution, lipoaspirate, bone marrow aspirate, a fraction thereof, a concentrate thereof, or a combination thereof.

11. The method according to claim 9 , wherein the blood-derived composition is platelet-rich plasma.

12. The method according to claim 9 , wherein step (a) consists of contacting whole blood, bone marrow aspirate, lipoaspirate, a fraction thereof, a concentrate thereof, or a combination thereof, with polyacrylamide beads to form a concentrated composition, and separating the concentrated composition from the polyacrylamide beads to produce the blood-derived composition.

13. The method according to claim 9 , wherein the concentration of IL-4, IL-10, IL-1ra, or TGFβ-1 in the blood-derived composition is greater than the respective concentration of IL-4, IL-10, IL-1ra, or TFBβ-1 in whole blood.

14. A method for mediating the phenotype of macrophages, consisting of:
(a) obtaining a composition comprising interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-1 receptor antagonist (IL-1ra), transforming growth factor β-1 (TGF-β-1), or a combination thereof, wherein the IL-4, IL-10, IL-1ra, TGF-β-1 is derived from blood, bone marrow aspirate, lipoaspirate, a fraction thereof, a concentrate thereof, or a combination thereof; and
(b) culturing an isolated source of macrophages in a activating composition consisting of the composition and optionally a non-differentiating growth media for a period of time, wherein the macrophages display a polarized phenotype after the period of time.

15. The method according to claim 14 , wherein the period of time is at least twelve hours.

16. The method according to claim 14 , wherein the period of time is about ten days.

17. The method according to claim 14 , wherein the composition comprises about 25% by volume of the culture medium.

18. The method according to claim 14 , wherein at least about 50% of the macrophages display an M2 phenotype after the period of time.

19. The method according to claim 14 , wherein step (a) includes contacting blood, bone marrow aspirate, lipoaspirate, a fraction thereof, a concentrate thereof, or a combination thereof, with a concentrating material to form an activating composition, and separating the activating composition from the concentrating material to form the blood-derived composition.

20. The method according to claim 14 , wherein the concentration of at least one of IL-4, IL-10, IL-1ra, or TGFβ-1 in the composition is greater than the respective concentration of IL-4, IL-10, IL-1ra, or TFBβ-1 in whole blood.

Assignees

• Biomet Biologics Llc
• Biomet Biologies Llc

Inventors

• Woodell-May Jennifer E
• Higgins Joel C
• King William

Classifications

• A61K35/16

  Blood plasma; Blood serum (umbilical cord blood A61K35/51) · CPC title

• C12N5/0667

  Adipose-derived stem cells [ADSC]; Adipose stromal stem cells · CPC title

• C12N2502/1382

  Adipose-derived stem cells [ADSC], adipose stromal stem cells · CPC title

• C12N5/0645 Primary

  Macrophages, e.g. Kuepfer cells in the liver; Monocytes · CPC title

• A61K35/15 Primary

  Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells (presenting a specific antigen A61K39/00; therapeutic combinations of antibodies, or fragments thereof, and blood-derived cells A61K39/00) · CPC title