Cell programming

Abstract

The present invention is concerned with methods for reprogramming of mammalian somatic cells and in particular to reprogramming of mature mammalian somatic cells into multi-potent precursor cells.

1 . A method of reprogramming a mature mammalian somatic cell into a reprogrammed multi-potent lineage-specific precursor cell, said method comprising the steps of:
 a) delivering one or more factors to said somatic cell, wherein said one or more factors determine the lineage specificity of said precursor cell; and   b) culturing said somatic cell under conditions permissive to the culture of said lineage-specific precursor cell.   
     
     
         2 . The method according to  claim 1  wherein said mature mammalian somatic cell is a mature mammalian fibroblast selected from lung fibroblasts, kidney fibroblasts, cardiac fibroblasts, stromal fibroblasts, foreskin fibroblasts or dermal fibroblasts. 
     
     
         3 . The method according to  claim 1  wherein said mature mammalian somatic cell is a mature human dermal fibroblast. 
     
     
         4 . The method according to  claim 1  wherein said mature mammalian somatic cell is a cell from a patient suffering from a neurological disorder or injury in which tissue regeneration is a component of healing and wherein said reprogrammed multi-potent lineage-specific precursor cell is a disease-specific reprogrammed multi-potent lineage-specific precursor cell. 
     
     
         5 . The method according to  claim 1  wherein said reprogrammed multi-potent, lineage-specific precursor cell is a multi-potent neural precursor cell. 
     
     
         6 . The method of  claim 5  wherein said multi-potent neural precursor cell expresses at least one neural cell lineage marker selected from the group consisting of Pax6, Sox2, Hes 1, Hes 5, Sox1, Sox3, Mash 1/Ashl 1 and neurogenin 2. 
     
     
         7 . The method according to  claim 1  wherein said step of delivering said one or more factors to said somatic cell includes the delivery of said one or more factors via protein transduction or via protein expression from non-viral or viral vectors. 
     
     
         8 . The method according to  claim 1  wherein said one or more factors are selected from proteins such as transcription factors, nucleic acids encoding said transcription factors, small molecules capable of influencing the amount of said transcription factors present in said somatic cell, or any combination thereof. 
     
     
         9 . The method according to  claim 8  wherein said transcription factors are the transcription factors Sox2 and Pax6 or any known transcription factors which, alone or in combination, are capable of producing multi-potent neural precursor cells. 
     
     
         10 . The method according to  claim 1  wherein said step of culturing said somatic cell includes culturing said cell in medium capable of supporting growth of said precursor cells. 
     
     
         11 . The method according to  claim 10  wherein said medium is supplemented with a chromatin modifying agent capable of facilitating reprogramming of said somatic cell and wherein said chromatin modifying agent is selected from agents promoting acetylation of chromatin, inhibiting deacetylation of chromatin, altering histone methylation states within chromatin or leading to DNA demethylation within chromatin. 
     
     
         12 . The method according to  claim 11  wherein said chromatin modifying agent is valproic acid at 1 μM. 
     
     
         13 . The method according to  claim 12  wherein the valproic acid is used at a concentration of 1 μM. 
     
     
         14 . A reprogrammed multi-potent lineage-specific precursor cell produced by a method according to  claim 1 . 
     
     
         15 . A reprogrammed multi-potent lineage-specific precursor cell according to  claim 14 , wherein said precursor cell is a multi-potent neural precursor cell. 
     
     
         16 . The method according to  claim 1  wherein said mature mammalian somatic cell is a mature human dermal fibroblast and wherein said mature human dermal fibroblast is from a patient suffering from a neurological disorder or injury in which tissue regeneration is a component of healing and wherein said reprogrammed multi-potent lineage-specific precursor cell is a disease-specific reprogrammed multi-potent lineage-specific precursor cell. 
     
     
         17 . The method according to  claim 1  wherein said mature mammalian somatic cell is a mature human dermal fibroblast and wherein said reprogrammed multi-potent, lineage-specific precursor cell is a multi-potent neural precursor cell. 
     
     
         18 . The method of  claim 19  wherein said multi-potent neural precursor cell expresses at least one neural cell lineage marker selected from the group consisting of Pax6, Sox2, Hes 1, Hes 5, Sox1, Sox3, Mash 1/Ashl 1 and neurogenin 2. 
     
     
         19 . The method according to  claim 1  wherein said mature mammalian somatic cell is a mature human dermal fibroblast and wherein said step of delivering said one or more factors to said somatic cell includes the delivery of said one or more factors via protein transduction or via protein expression from non-viral or viral vectors. 
     
     
         20 . The method according to  claim 1  wherein said mature mammalian somatic cell is a mature human dermal fibroblast and wherein said one or more factors are selected from proteins such as transcription factors, nucleic acids encoding said transcription factors, small molecules capable of influencing the amount of said transcription factors present in said somatic cell, or any combination thereof.