Directed in vitro differentiation of marrow stromal cells into neural cell progenitors

Abstract

The invention relates to methods for inducing marrow stromal cells to differentiate into neural cells by way of increasing intracellular levels of cyclic AMP. The invention also encompasses methods of producing a neural cell by causing a marrow stromal cell to differentiate into a neural cell by increasing intracellular levels of cyclic AMP. Methods for treating a human patient in need of neural cells are also disclosed, as well as methods for treating a human patient having a disease, condition, or disorder of the central nervous system.

We claim:  
     
         1 . A method of producing a neural cell, said method comprising contacting an isolated marrow stromal cell with a cyclic AMP-stimulating compound, thereby increasing the intracellular level of cyclic AMP in said marrow stromal cell and producing a neural cell.  
     
     
         2 . The method of  claim 1 , wherein said cyclic AMP-stimulating compound is selected from the group consisting of epinephrine, isoproterenol, forskolin, isobutylmethylxanthine (IBMX), dibutyryl cyclic AMP (dbcAMP), and a combination of IBMX and dbcAMP.  
     
     
         3 . The method of  claim 2 , wherein said cyclic AMP-stimulating compound is a combination of IBMX and dbcAMP.  
     
     
         4 . The method of  claim 3 , wherein said IBMX is present in a concentration of from about 0.01 millimolar to about 5.0 millimolar, and wherein said dbcAMP is present in a concentration of from about 0.1 millimolar to about 10.0 millimolar.  
     
     
         5 . The method of  claim 4 , wherein said IBMX is present at 0.5 millimolar and further wherein said dbcAMP is present at 1 millimolar.  
     
     
         6 . The neural cell of  claim 1 , wherein said cell is human.  
     
     
         7 . A method of treating a human patient having a disease, disorder, or condition of the central nervous system, said method comprising administering to a patient neural cells produced by the method of  claim 1 , thereby treating the disease, disorder, or condition.  
     
     
         8 . The method of  claim 7 , wherein said neural cells are transfected with an isolated nucleic acid encoding a therapeutic protein, wherein when said protein is expressed in said cells said protein serves to effect treatment of said disease, disorder, or condition.  
     
     
         9 . The neural cell of  claim 8 , wherein said cell is human.  
     
     
         10 . The method of  claim 7 , wherein said disease, disorder, or condition of the central nervous system is selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, a tumor, a trauma, elderly dementia, Tay-Sach's disease, Sandhoff s disease, Hurler's syndrome, Krabbe's disease, birth-induced traumatic central nervous system injury, epilepsy, multiple sclerosis, trauma, tumor, stroke, and spinal cord injury.  
     
     
         11 . A method of treating a human patient in need of neural cells, said method comprising obtaining marrow stromal cells from a human donor, producing neural cells by the method of  claim 1 , and transplanting said neural cells into said human patient in need of said neural cells, thereby treating said human patient in need of neural cells.  
     
     
         12 . A method of inducing differentiation of an isolated marrow stromal cell into a neural cell, said method comprising contacting said marrow stromal cell with a cyclic AMP stimulating compound.  
     
     
         13 . The method of  claim 12 , wherein said compound is selected from the group consisting of epinephrine, isoproterenol, forskolin, IBMX, dbcAMP, and a combination of IBMX and dbcAMP.  
     
     
         14 . The method  claim 13 , wherein said compound is a combination of IBMX and dbcAMP.  
     
     
         15 . The method of  claim 14 , wherein said IBMX is present in a concentration of from about 0.01 millimolar to about 5.0 millimolar and wherein said dbcAMP is present in a concentration of from about 0.01 millimolar to about 10.0 millimolar.  
     
     
         16 . The method of  claim 15 , wherein said IBMX is present at a concentration of 0.5 millimolar and wherein said dbcAMP is present at a concentration of 1 millimolar.  
     
     
         17 . The neural cell of  claim 9 , wherein said cell is human.  
     
     
         18 . A method of treating a human patient having a disease, disorder, or condition of the central nervous system, said method comprising administering to a patient neural cells produced by the method of  claim 12 .  
     
     
         19 . The method of  claim 18 , wherein said neural cells are transfected with an isolated nucleic acid encoding a therapeutic protein, wherein when said protein is expressed in said cells said protein serves to effect treatment of said disease, disorder, or condition.  
     
     
         20 . The neural cell of  claim 19 , wherein said cell is human.