Method for treating a condition with neural progenitor cells derived from whole bone marrow

Abstract

A method is described for generating a clinically significant volume of neural progenitor cells from whole bone marrow. A mass of bone marrow cells may be grown in a culture supplemented with fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF). Further methods of the present invention are directed to utilizing the neural progenitor cells cultured in this fashion in the treatment of various neuropathological conditions, and in targeting delivery of cells transfected with a particular gene to diseased or damaged tissue.

1 - 15 . (canceled)  
     
     
         16 . A method to treat a neuropathologic condition, the method comprising: 
 providing a mass of cells obtained from the whole bone marrow of a mammal including at least one stem cell; and    culturing the mass of cells in a medium comprising fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) to produce a neural progenitor cell; and    administering the neural progenitor cell to a mammal.    
     
     
         17 . (canceled)  
     
     
         18 . The method of  claim 16 , wherein the mass of cells is obtained from an adult mammal.  
     
     
         19 . The method of  claim 16 , wherein the mass of cells is obtained from a fetus.  
     
     
         20 . (canceled)  
     
     
         21 . The method of  claim 16 , wherein the medium is Dulbecco's modified Eagle medium (DMEM).  
     
     
         22 . The method of  claim 16 , wherein the medium further comprises a supplement.  
     
     
         23 . The method of  claim 22 , wherein the supplement is selected from the group consisting of B27, N2, and combinations thereof.  
     
     
         24 . The method of  claim 16 , wherein the medium further comprises an additional compound.  
     
     
         25 . The method of  claim 24 , wherein the additional compound is selected from the group consisting of interleukin-3 (IL-3), stem cell factor-1 (SCF-1), sonic hedgehog (Shh), fms-like tyrosine kinase-3 (Flt3) ligand, leukemia inhibitory factor (LIF), and combinations thereof.  
     
     
         26 . The method of  claim 16 , wherein the medium further comprises an antibiotic.  
     
     
         27 . The method of  claim 26 , wherein the antibiotic is selected from the group consisting of penicillin, streptomycin, and combinations thereof.  
     
     
         28 . The method of  claim 16 , wherein clusters of neural progenitor cells develop in the medium, and the method further comprises separating at least one cluster from the medium.  
     
     
         29 . The method of  claim 28 , further including subculturing the at least one cluster.  
     
     
         30 . The method of  claim 29 , wherein subculturing the at least one cluster further comprises culturing the at least one cluster in a medium comprising FGF-2 and EGF.  
     
     
         31 . The method of  claim 16 , wherein the neuropathologic condition is selected from the group consisting of brain tumor, brain ischemia, stroke, spinal cord injury, and neurodegenerative disorders.  
     
     
         32 . The method of  claim 16 , wherein administering the neural progenitor cell further comprises administering a clinically significant volume of neural progenitor cells.  
     
     
         33 . The method of  claim 32 , wherein the clinically significant volume of neural progenitor cells is sufficient to treat the neuropathologic condition.  
     
     
         34 - 52 . (canceled)  
     
     
         53 . A method to treat a neuropathologic condition, the method comprising: 
 providing a clinically significant volume of neural progenitor cells, produced by a process, comprising: 
 providing a mass of cells obtained from the whole bone marrow of a mammal including at least one stem cell, and  
 culturing the mass of cells in a medium comprising fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) to produce the clinically significant volume of neural progenitor cells; and  
   administering said clinically significant volume of neural progenitor cells to a mammal to treat said neuropathologic condition.    
     
     
         54 . The clinically significant volume of neural progenitor cells of  claim 53 , wherein the medium further comprises a supplement selected from the group consisting of B27, N2, and combinations thereof.  
     
     
         55 . The clinically significant volume of neural progenitor cells of  claim 53 , wherein the medium further comprises an additional compound selected from the group consisting of interleukin-3 (IL-3), stem cell factor-1 (SCF-1), sonic hedgehog (Shh), fms-like tyrosine kinase-3 (Flt3) ligand, leukemia inhibitory factor (LIF), and combinations thereof.  
     
     
         56 . The clinically significant volume of neural progenitor cells of  claim 53 , wherein clusters of the neural progenitor cells develop in the medium, and the process further comprises: 
 separating at least one cluster from the medium; and    subculturing the at least one cluster in a medium comprising FGF-2 and EGF.    
     
     
         57 . A method to treat a neuropathologic condition, the method comprising: 
 providing a mass of cells obtained from the whole bone marrow of a mammal including at least one stem cell;    culturing the mass of cells in Dulbecco's modified Eagle Medium (DMEM) comprising B27, penicillin, streptomycin, fibroblast growth factor-2 (FGF-2), epidermal growth factor (EGF) and an additional compound, whereby sphere clusters of the neural progenitor cells develop in the medium;    separating at least one sphere cluster from the medium;    subculturing the at least one sphere cluster in a medium comprising FGF-2 and EGF to produce a clinically significant volume of neural progenitor cells; and    administering said clinically significant volume of neural progenitor cells to a mammal to treat said neuropathologic condition.    
     
     
         58 . A method to treat a neuropathologic condition, the method comprising: 
 providing a clinically significant volume of neural progenitor cells, produced by a process, comprising: 
 providing a mass of cells obtained from the whole bone marrow of a mammal including at least one stem cell,  
 culturing the mass of cells in Dulbecco's modified Eagle Medium (DMEM) comprising B27, penicillin, streptomycin, fibroblast growth factor-2 (FGF-2), and epidermal growth factor (EGF), whereby sphere clusters of the neural progenitor cells develop in the medium,  
 separating at least one sphere cluster from the medium, and  
 subculturing the at least one sphere cluster in a medium comprising FGF-2 and EGF to produce a clinically significant volume of neural progenitor cells; and  
   administering the neural progenitor cells to a mammal to treat said neuropathologic condition.