Methods for making cytokine compositions from tissues using non-centrifugal methods
AssigneeBiomet Manufacturing, LLC
InventorsMichael D. Leach, Jennifer E. Woodell-May, Joel C. Higgins, Krista O'Shaughnessey
Life of this patent
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Abstract
Non-centrifugal methods for generating a solution rich in interleukin-1 receptor antagonist from a tissue comprising cytokine-producing cells. The solution rich in IL-1ra can also include at least one of sTNF-RI, sTNF-RII, IGF-I, EGF, HGF, PDGF-AB, PDGF-BB, VEGF, TGF-β1, and sIL-1 RII.
Claims26
A method for generating a solution rich in interleukin-1 receptor antagonist (IL-1ra) comprising:
- obtaining a liquid volume comprising white blood cells by separating white blood cells from a tissue comprising any one of:
- (a) whole blood, bone marrow aspirate, adipose tissue;
- (b) a fraction thereof;
- or (c) a mixture of any of the foregoing, using dielectrophoretic field-flow-fractionation (DEP-FFF), wherein separating the white blood cells concentrates the white blood cells to at least about 15,000/μl;
- and
- contacting the liquid volume comprising white blood cells with a solid extraction material to obtain a solution rich in IL-1ra.
A method of preparing an enriched biological solution, comprising:
- (a) separating white blood cells from a tissue selected from whole blood, bone marrow aspirate, adipose tissue, a fraction thereof, or a mixture thereof to produce a volume of concentrated white blood cells, including subjecting the tissue to di electrophoretic field-flow-fractionation (DEP-FFF), wherein the separating separates at least about one-half of red blood cells in the tissue from the volume of white blood cells;
- (b) mixing the volume of white blood cells with a liquid medium to form a cytokine cell suspension, wherein the cytokine cell suspension comprises white blood cells, and at least one of platelets, adipose stromal cells, and mesenchymal stromal cells;
- (c) contacting the cytokine cell suspension with polyacrylamide beads for a period of time between about 30 seconds and about 72 hours;
- and
- (d) separating liquid from the polyacrylamide beads to obtain a solution rich in IL-1ra and rich in at least one of epidermal growth factor (EGF), hepatocyte growth factor (HGF), platelet-derived growth factor-BB (PDGF-BB), and soluble interleukin-1 receptor II (sIL-1RII).
A method for generating a biological solution enriched in anti-inflammatory cytokines and growth factors, the method comprising:
- subjecting a tissue comprising any one of:
- (a) whole blood, bone marrow aspirate, adipose tissue;
- (b) a fraction thereof;
- or (c) a combination of any of the foregoing, to dielectrophoretic field-flow-fractionation to separate white blood cells from the tissue to produce a volume of white blood cells at a concentration of at least about 15,000/μl, wherein the volume of white blood cells includes about one-half or less of red blood cells from the tissue;
- and
- contacting the volume of white blood cells with a solid extraction material to obtain the biological solution enriched in at least one protein selected from interleukin-1 receptor II, platelet-derived growth factor BB, epidermal growth factor, and hepatocyte growth factor.
Dependencies are inferred from the claim text.