Multipotent neural stemcells from peripheral tissues and uses thereof

Abstract

This invention relates to multipotent neural stem cells, purified from the peripheral nervous system of mammals, capable of differentiating into neural and non-neural cell types. These stem cells provide an accessible source for autologous transplantation into CNS, PNS, and other damaged tissues.

1 . A multipotent stem cell substantially purified from skin of a mammal.  
     
     
         2 . The multipotent stem cell of  claim 1 , wherein said skin comprises the dermal layer.  
     
     
         3 . The multipotent stem cell of  claim 1 , wherein said skin is from a postnatal mammal.  
     
     
         4 . The multipotent stem cell of  claim 1 , wherein said skin is from an adult mammal.  
     
     
         5 . The multipotent stem cell of  claim 1 , wherein said skin is from a juvenile mammal.  
     
     
         6 . The multipotent stem cell of  claim 1 , wherein said mammal is a human.  
     
     
         7 . The multipotent stem cell of  claim 1 , wherein said cell expresses nestin.  
     
     
         8 . The multipotent stem cell of  claim 1 , wherein said cell expresses fibronectin.  
     
     
         9 . The multipotent stem cell of  claim 1 , which under appropriate conditions can be differentiated into a neuron, an astrocyte, a Schwann cell, or an oligodendrocyte.  
     
     
         10 . The multipotent stem cell of  claim 1 , which can be differentiated into a non-neural cell.  
     
     
         11 . The multipotent stem cell of  claim 9 , wherein said non-neural cell is a smooth muscle cell or an adipocyte.  
     
     
         12 . The multipotent stem cell of  claim 1 , wherein said cell contains a heterologous gene in an expressible genetic construction.  
     
     
         13 . The multipotent stem cell of  claim 11 , wherein said gene encodes a therapeutic protein.  
     
     
         14 . The multipotent stem cell of  claim 11 , wherein said gene encodes a cell fate-determining protein.  
     
     
         15 . A population of at least ten cells, wherein at least 30% of the cells are multipotent stem cells substantially purified from skin of a mammal.  
     
     
         16 . The population of cells of  claim 15 , wherein said skin is from a human.  
     
     
         17 . A composition comprising (i) a multipotent stem cell substantially purified from skin of a mammal, and (ii) a pharmaceutically acceptable carrier, auxiliary or excipient.  
     
     
         18 . A cell aggregate comprising at least ten multipotent stem cells derived from skin of a mammal, said multipotent stem cells capable of differentiating as dopaminergic neurons.  
     
     
         19 . The aggregate of  claim 17 , wherein said cells grow detached from the culture substrate.  
     
     
         20 . The aggregate of  claim 18 , wherein said aggregate comprises at least one hundred multipotent neural stem cells derived from skin of a mammal.  
     
     
         21 . A method of producing a population of at least ten cells, wherein at least 30% of the cells are multipotent stem cells substantially purified from skin of a postnatal mammal, or progeny of said multipotent stem cells, said method comprising: 
 (a) providing said skin from said mammal;    (b) culturing said skin under conditions in which multipotent stem cells proliferate and in which at least 25% of the cells that are not multipotent stem cells die or attach to the culture substrate; and    (c) continuing culture step (b) until at least 30% of the cells are multipotent stem cells or progeny of said multipotent stem cells.    
     
     
         22 . A method of producing a population of at least ten cells, wherein at least 30% of the cells are multipotent stem cells substantially purified from skin of a postnatal mammal or progeny of said multipotent stem cells, said method comprising: 
 (a) providing said skin from said mammal;    (b) culturing said skin under conditions in which multipotent stem cells proliferate and in which at least 25% of the cells that are not multipotent stem cells die or attach to the culture substrate;    (c) separating said multipotent stem cells from said cells that attach to said culture substrate; and    (d) repeating steps (b) and (c) until at least 30% of the cells are multipotent stem cells or progeny of said multipotent stem cells.    
     
     
         23 . The method of  claim 22 , wherein said population is at least one hundred cells.  
     
     
         24 . A multipotent stem cell in the central nervous system of a mammal, said multipotent stem cell produced by a method of comprising transplanting a multipotent stem cell substantially purified from the skin of a mammal into the central nervous system of said mammal.  
     
     
         25 . The multipotent stem cell of  claim 24 , wherein said mammal from which said cell is substantially purified is said mammal into which said cell is transplanted.  
     
     
         26 . The multipotent stem cell of  claim 25 , wherein said mammal from which said cell is substantially purified is immunologically similar to said mammal into which said cell is transplanted.  
     
     
         27 . A kit comprising a multipotent stem cell substantially purified from skin of a mammal.  
     
     
         28 . The kit of  claim 27 , said kit comprising a population of cells, wherein at least 30% of said cells are multipotent stem cells substantially purified from said skin.  
     
     
         29 . A kit for the substantial purification of multipotent stem cells from skin of a mammal.  
     
     
         30 . A method of treating a patient having a disease characterized by failure of a cell type, said method comprising administering to said patient a multipotent stem cell derived from a peripheral tissue, said multipotent stem cell capable of producing neurons and glia.  
     
     
         31 . The method of  claim 30 , wherein said peripheral tissue is skin.  
     
     
         32 . A kit for use in performing the method of  claim 30 .  
     
     
         33 . A method for treating a patient having a disease characterized by failure of a cell type, said method comprising admininstering to said patient a differentiated cell that is the progeny of a multipotent stem cell derived from skin.  
     
     
         34 . The method of  claim 33 , wherein said skin is from a postnatal mammal.  
     
     
         35 . A method for making a neuron that expresses dopamine, said method comprising: 
 (a) providing a multipotent stem cell derived from skin; and    (b) culturing said cell under conditions whereby said cell differentiates into a neuron that expresses dopamine.    
     
     
         36 . A method for making a cell other than a skin cell, said method comprising culturing a multipotent stem cell substantially purified from peripheral tissue of a mammal under conditions in which said cell differentiates as a cell other than a skin cell.  
     
     
         37 . The method of  claim 36 , wherein said cell is a neuron, a glial cell, a cardiac cell, a pancreatic islet cell, an adipocyte or a smooth muscle cell.  
     
     
         38 . The method of  claim 36 , wherein said peripheral tissue is skin.  
     
     
         39 . A substantially pure composition of multipotent stem cells, which multipotent stem cell differentiates into neural cell types and non-neural cell types.  
     
     
         40 . The stem cells of  claim 39 , wherein said non-neural cell type is an adipocyte, a smooth muscle cell or a cardiac cell.  
     
     
         41 . An ex vivo cellular composition wherein at least 30% of the cells are progeny of the multipotent stem cells of  claim 39 .  
     
     
         42 . A cellular composition of  claim 41 , wherein said progeny is undifferentiated progeny.  
     
     
         43 . A cellular composition of  claim 42 , wherein said progeny is capable of differentiating into a neural or non-neural cell.