Stem Cells from the Mammalian Neural Plate

Abstract

The present invention relates to methods for deriving novel stem cells from the mammalian early neural plate.

1 . (canceled) 
     
     
         2 . (canceled) 
     
     
         3 . A method of maintaining a neural plate stem cell (NPSC) in culture, comprising:
 (a) providing a neural plate stem cell; and   culturing the cell population in the presence of FGF4   
       wherein FGF4 maintains the NPSC in a pre-patterning state and FGF2 does not maintains the NPSC in a pre-patterning state. 
     
     
         4 . The method of  claim 3  wherein the “pre-patterning state” is a state where the NPSC expresses the marker Brn-2 and does not express:
 (i) the neurogenic bHLH factors Ngn2 and MASH1; 
 (ii) MASH1; 
 (iii) Pax3; 
 (iv) Pax6; 
 (v) En1; 
 (vi) En2; and/or 
 (vii) Krox20. 
 
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         7 . (canceled) 
     
     
         8 . The method of  claim 7  wherein the pluripotent cell or primitive neurectodermal cell is cultured in the presence of only FGF4. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 3 , wherein the neural plate stem cell expresses brn2. 
     
     
         11 . The method of  claim 3 , wherein the neural plate stem cell does not express one or more of the markers selected from the group of PLZF, ngn2 or MASH1. 
     
     
         12 . The method of  claim 3 , wherein the cell population is cultured in the absence of fibronectin. 
     
     
         13 . An isolated neural plate stem cell, or population of such cells maintained in culture for at least 12 passages in the presence of FGF4, wherein the NPSC maintain expression of the marker Brn-2 and do not express the markers Ngn2, MASH!, 50 Pax3, Pax6, En1, En2, and Krox20 on the NPSC, 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . An isolated neural plate stem cell, characterised in that:
 (i) FGF4 increases the proliferation of the NPSC; and   (ii) FGF2 does not increase the proliferation of the neural plate stem cell.   
     
     
         17 . The isolated neural plate stem cell of  claim 16  further characterized by expression of brn2. 
     
     
         18 . The isolated neural plate stem cell of  claim 17  further characterized by the absence of expression of:
 (i) Ngn2 and MASH1; 
 (ii) MASH1; 
 (iii) Pax3; 
 (iv) Pax6; 
 (v) En1; 
 (vi) En2; and/or 
 (vii) Krox20. 
 
     
     
         19 . The isolated population of neural plate stem cells of  claim 18 , characterized in that;
 (i) substantially all of the cells in the population express brn2.   
     
     
         20 . The isolated population of neural plate stem cells of  claim 19 , wherein fewer than 95% of the cells in the population express sox1. 
     
     
         21 . The isolated population of neural plate stem cells of  claim 20  wherein fewer than 90% of the NPSC population expresses Sox-1, for example less than 85%, 75%, 65%, 55%, 45%, 35%, 25%, 15% or less than 5%. 
     
     
         22 . The isolated population of neural plate stem cells of  claim 19 , characterized by the absence of expression of:
 (i) Ngn2 and MASH1;   (ii) MASH1;   (iii) Pax3;   (iv) Pax6;   (v) En1;   (vi) En2; and/or   (vii) Krox20.   
     
     
         23 . The isolated population of neural plate stem cells of  claim 19 , wherein the cell population proliferates in the presence of FGF4 and does not proliferate in the presence of FGF2. 
     
     
         24 . (canceled) 
     
     
         25 . The isolated neural plate stem cell of  claim 23 , which is capable of differentiation into a monoaminergic neuron when cultured in the presence of Shh and FGF8 and the absence of FGF4. 
     
     
         26 . The isolated neural plate stem cell of  claim 23 , which is capable of differentiation into motor neuron when cultured in the presence of retinoic acid the absence of FGF4. 
     
     
         27 . The isolated neural plate stem cell of  claim 23 , which is capable of differentiation into any neural or glial cell type. 
     
     
         28 . The isolated neural plate stem cell population of  claim 23  capable of differentiation into monoaminergic neuron or motor neuron after at least 40 passages.