Induction Medium & Methods for Stem Cell Culture & Therapy

Abstract

Novel MSC stem-cell culture and therapy methods and culture medium compositions for the purpose of inducing, activating, or priming discrete uniform cell phenotypes to selectively promote or suppress inflammation and immunity, yielding primed, activated, or induced cells used in cell-based therapy.

I claim: 
     
         1 . An induction medium for inducing a cultured population of MSC stem cells, comprising:
 a Toll-like receptor (TLR) ligand or TLR-ligand inducer;   erythropoietin (EPO);   exposure to hypoxia or hypoxia mimetic; and   other known, standard culture-medium components;   where said TLR ligand induces said MSC stem cells in a predicable manner, yielding expanded and induced, primed, or activated populations of MSC stem cells.   
     
     
         2 . The induction medium of  claim 1 , where said TLR ligand amount is between about 0.10 picomolar (pM) and about 100 millimolar (mM), or equivalent TLR-ligand inducer amount; said erythropoietin (EPO) amount is between about 0.5 mU/mL and about 50 mU/mL; said exposure to hypoxia is about 0.5% to about 2% oxygen conditions; and said hypoxia mimetic further comprises cobalt chloride or desferrioxamine, at a concentration of about 10 uM to about 1 mM. 
     
     
         3 . The induction medium of  claim 1 , where said TLR ligand further comprises, exclusively or in various combinations, 1L4, IL13, poly(A:U), poly(I:C), aminoalkyl glucosaminide 4-phosphates, interferons, TNF-alpha, GM-CSF, and lipopolysaccharide (IPS). 
     
     
         4 . The induction medium of  claim 1 , where said TLR ligand further comprises, exclusively or in various combinations, 1L4, IL13, poly(A:U), poly(I:C), or substitutes thereof. 
     
     
         5 . The induction medium of  claim 1 , where said TLR ligand further comprises, exclusively or in various combinations, aminoalkyl glucosaminide 4-phosphates, interferons, TNF-alpha, GM-CSF, lipopolysaccharide (LPS), or substitutes thereof. 
     
     
         6 . The culture medium of  claim 1 , where said TLR ligand or TLR-ligand inducer, said erythropoietin (EPO), and said exposure to hypoxia or hypoxia mimetic is provided as a culture-medium induction supplement to be added to various other known, standard culture-medium components having various desired characteristics. 
     
     
         7 . The culture medium of  claim 1 , further comprising a hermetically-sealed culture vessel having a solid surface; and containing said induction medium in full or supplement form. 
     
     
         8 . A method for inducing, activating, or priming MSC stein cells, comprising:
 providing a population of MSC stem cells;   providing an induction medium comprising:
 a Toll-like receptor (TLR) ligand or TLR-ligand inducer; 
 erythropoietin (EPO); 
 exposure to hypoxia or hypoxia mimetic; and 
 other known, standard culture-medium components; 
 where said TLR ligand induces said MSC stem cells in a predicable manner, yielding expanded and induced, primed, or activated populations of MSC stem cells; 
   contacting said MSC stem cells with the induction medium; and   culturing said MSC stem cells under appropriate conditions.   
     
     
         9 . The method for inducing, activating, or priming MSC stem cells of  claim 8 , where said TLR ligand amount is between about 0.10 picomolar (pM) and about 100 millimolar (mM), or equivalent TLR-ligand inducer amount; said erythropoietin (EPO) amount is between about 0.5 mU/mL and about 50 mU/mL; said exposure to hypoxia is about 0.5% to about 2% oxygen conditions; and said hypoxia mimetic further comprises cobalt chloride or desferrioxamine, at a concentration of about 10 uM to about 1 mM. 
     
     
         10 . method for inducing, activating, or priming MSC stem cells of  claim 8 , where said TLR ligand further comprises, exclusively or in various combinations, IL4, IL13, poly(A:U), poly(I:C), aminoalkyl glucosaminide 4-phosphates, interferons, TNF-alpha, GM-CSF, and lipopolysaccharide (LPS). 
     
     
         11 . The method for inducing, activating, or priming MSC stem cells of  claim 8 , where said TLR ligand further comprises, exclusively or in various combinations, IL4, IL13, poly(A:U), poly(I:C), or substitutes thereof. 
     
     
         12 . The method for inducing, activating, or priming MSC stein cells of  claim 8 , where said TLR ligand further comprises, exclusively or in various combinations, aminoalkyl glucosaminide 4-phosphates, interferons, TNF-alpha, GM-CSF, lipopolysaccharide (LPS), or substitutes thereof. 
     
     
         13 . A method of cellular therapy, comprising:
 ex-vivo culturing of expanded and induced, primed, or activated populations of MSC stem cells in an appropriate induction medium comprising:
 a Toll-like receptor (TLR) ligand or TLR-ligand inducer; 
 erythropoietin (EPO); 
 exposure to hypoxia or hypoxia mimetic; and 
 other known, standard, variable culture-medium components; 
 where said TLR ligand induces said MSC stem cells in a predicable manner, yielding 
 expanded and induced, primed, or activated populations of MSC stem cells; and 
   using the yielded expanded and induced, primed, or activated populations of MSC stem cells as a medicament or therapeutic agent fir cellular therapy.   
     
     
         14 . The method of cellular therapy of  claim 13 , where said TLR ligand amount is between about 0.10 picomolar (pM) and about 100 millimolar (mM), or equivalent TLR-ligand inducer amount; said erythropoietin (EPO) amount is between about 0.5 mU/mL, and about 50 mU/mL; said exposure to hypoxia is about 0.5% to about 2% oxygen conditions;
 and said hypoxia mimetic further comprises cobalt chloride or desferrioxamine, at a concentration of about 10 uM to about 1 mM.   
     
     
         15 . The method of cellular therapy of  claim 13 , where said TLR ligand further comprises, exclusively or in various combinations, IL4, IL13, poly(A:U), poly(I:C), aminoalkyl glucosaminide 4-phosphates, interferons, TNF-alpha, GM-CSF, and lipopolysaccharide (LPS). 
     
     
         16 . The method of cellular therapy of  claim 13 , where said TLR ligand further comprises, exclusively or in various combinations, IL4, IL13, poly(A:U), poly(I:C), or substitutes thereof. 
     
     
         17 . The method of cellular therapy of  claim 13 , where said TLR ligand further comprises, exclusively or in various combinations, aminoalkyl glucosaminide 4-phosphates, interferons, TNF-alpha, GM-CSF, lipopolysaccharide (LPS), or substitutes thereof.