Bioactive compositions derivable from platelet concentrates, and methods for preparing and using same

Abstract

The present disclosure provides a composition comprising a bioactive fraction derived from a platelet concentrate, methods of making the bioactive fraction, and culture medium supplemented with the bioactive fraction. Preferred bioactive fractions have relatively low fibrinogen concentrations while retaining native growth factors in beneficial amounts and ratios.

1 - 25 . (canceled) 
     
     
         26 . A method for preparing a bioactive composition, comprising:
 lysing platelets of a human blood-derived concentrate containing platelets and plasma, to form a lysed platelet preparation;   forming a gel clot from the lysed platelet preparation by converting fibrinogen to fibrin in the lysed platelet preparation;   compressing the gel clot to express liquid from the gel clot; and   separating the liquid from solids of the gel clot.   
     
     
         27 . A method for processing a platelet lysate composition, comprising:
 lysing platelets of a human blood-derived concentrate containing platelets and plasma, to form a lysed platelet preparation;   compressing a gel clot formed by converting fibrinogen to fibrin in a lysed platelet preparation so as to express liquid from the gel clot.   
     
     
         28 . A method for processing a platelet lysate composition, comprising:
 passing a liquid bioactive fraction of a human blood-derived platelet concentrate comprising native components of the platelet concentrate including fibrinogen, albumin, globulin, TGF-β1, EGF, FGF-beta, PDGF-AA, PDGF-BB, SDF-1α, and VEGF, wherein the fibrinogen is present at a level less than 20,000 ng/mL, through at least a first depth filter, so as to remove suspended solids from the liquid bioactive fraction.   
     
     
         29 . The method of  claim 28 , also comprising obtaining the liquid bioactive fraction from the blood-derived platelet concentrate without a centrifugation step to separate a clotted fraction of the concentrate from the liquid bioactive fraction of the concentrate. 
     
     
         30 . The method of  claim 28 , wherein said obtaining includes:
 lysing platelets of the concentrate to form a lysed preparation;   converting fibrinogen of the lysed preparation to fibrin so as to form a gel clot; and   compressing the gel clot without centrifugation so as to express the liquid bioactive fraction from the gel clot.   
     
     
         31 . The method of  claim 30 , wherein the compressing comprises compressing the gel clot between and in contact with a first surface and a second surface so as to express the liquid bioactive fraction from the gel clot and leave behind a solid clot mass. 
     
     
         32 . The method of any one of  claims 28 - 31 , wherein said passing results in:
 at least a 20% reduction in the level of one, some or all of TGF-beta-1, EGF, FGF-b, PDGF-AA, PDGF-BB, SDF-1α, and VEGF; and/or   at least a 50% reduction in the level of TGF-beta-1; and/or   at least a 30% reduction in the level of EGF; and/or   at least a 20% reduction in the level of FGF-b; and/or   at least a 50% reduction in the level of PDGF-AA; and/or   at least a 50% reduction in the level of PDGF-BB; and/or   at least a 20% reduction in the level of SDF-1α; and/or   at least a 30% reduction in the level of VEGF.   
     
     
         33 - 38 . (canceled) 
     
     
         39 . The method of  claim 26 , wherein the compressing comprises compressing the gel clot between and in contact with a first surface and a second surface so as to express the liquid bioactive fraction from the gel clot and leave behind a solid clot mass. 
     
     
         40 . The method of  claim 26 , wherein after said separating the liquid has a reduced concentration of fibrinogen as compared to that of the lysed platelet preparation prior to formation of the gel clot. 
     
     
         41 . The method of  claim 40 , wherein after said separating the liquid contains less than 5% of the fibrinogen present in the lysed platelet preparation prior to formation of the gel clot. 
     
     
         42 . The method of  claim 40 , wherein after said separating the liquid contains less than 2% of the fibrinogen present in the lysed platelet preparation prior to formation of the gel clot. 
     
     
         43 . The method of  claim 26 , wherein after said separating the liquid has a fibrinogen content of less than 20,000 ng/mL. 
     
     
         44 . The method of  claim 26 , wherein after said separating the liquid retains one or more plasma components including:
 0.5 to 2.5 g/dL globulins;   2 to 5 g/dL albumin;   100 to 200 mmol/L sodium;   40 to 70 mg/dL triglycerides; and/or   150 to 300 mg/dL glucose.   
     
     
         45 . The method of  claim 26 , wherein after said separating the liquid retains one or more growth factors including: TGF-β1, EGF, FGF-basic, PDGF-AA, PDGF-BB, SDF-1α, and/or VEGF. 
     
     
         46 . The method of  claim 27 , wherein the compressing comprises compressing the gel clot between and in contact with a first surface and a second surface so as to express a liquid bioactive fraction from the gel clot and leave behind a solid clot mass. 
     
     
         47 . The method of  claim 27 , wherein after said compressing the expressed liquid has a reduced concentration of fibrinogen as compared to that of the lysed platelet preparation prior to formation of the gel clot. 
     
     
         48 . The method of  claim 47 , wherein after said compressing the liquid contains less than 5% of the fibrinogen present in the lysed platelet preparation prior to formation of the gel clot. 
     
     
         49 . The method of  claim 47 , wherein after said separating the liquid contains less than 2% of the fibrinogen present in the lysed platelet preparation prior to formation of the gel clot. 
     
     
         50 . The method of  claim 47 , wherein after said compressing the liquid has a fibrinogen content of less than 20,000 ng/mL. 
     
     
         51 . The method of  claim 47 , wherein after said compressing the liquid retains one or more plasma components including:
 0.5 to 2.5 g/dL globulins;   2 to 5 g/dL albumin;   100 to 200 mmol/L sodium;   40 to 70 mg/dL triglycerides; and/or   150 to 300 mg/dL glucose.   
     
     
         52 . The method of  claim 27 , wherein after said compressing the liquid retains one or more growth factors including: TGF-β1, EGF, FGF-basic, PDGF-AA, PDGF-BB, SDF-1α, and/or VEGF. 
     
     
         53 . A method for preparing a bioactive composition, the method comprising:
 compressing a clotted platelet lysate material to separate a liquid portion from a solid clotted portion.   
     
     
         54 . The method of  claim 53 , wherein said solid clotted portion comprises a fibrin matrix, and wherein said compressing condenses said fibrin matrix. 
     
     
         55 . The method of  claim 53 , wherein said clotted platelet lysate material comprises a lysed platelet preparation which has been treated to convert fibrinogen to fibrin. 
     
     
         56 . The method of  claim 53 , wherein said liquid portion has a fibrinogen content of less than 20,000 ng/mL. 
     
     
         57 . The method of  claim 53 , wherein said liquid portion retains one or more plasma components from the clotted platelet lysate material including:
 0.5 to 2.5 g/dL globulins;   2 to 5 g/dL albumin;   100 to 200 mmol/L sodium;   40 to 70 mg/dL triglycerides; and/or   150 to 300 mg/dL glucose.   
     
     
         58 . The method of  claim 53 , wherein said liquid portion retains one or more growth factors from the clotted platelet lysate material including: TGF-β1, EGF, FGF-basic, PDGF-AA, PDGF-BB, SDF-1α, and/or VEGF. 
     
     
         59 . The method of  claim 55 , wherein said liquid portion has a reduced concentration of fibrinogen as compared to that of lysed platelet preparation prior to formation of the clotted platelet lysate material.