Method for discovering neurogenic agents

Abstract

A cell line and an assay for discovering neurogenic drugs are described. The assay allows for systematic screening of test agents such as libraries of compounds.

1 . A stable human neural progenitor cell line, wherein
 (i) the cell line is derived from the hippocampus;   (ii) the cell line has been expanded for at least ten cell doublings without substantially differentiating; and   (iii) the cell line is capable of generating at least 40% neurons upon differentiation by mitogen withdrawal.   
     
     
         2 . The cell line of  claim 1 , which is obtained by genetic modification to enhance the mitotic capacity of the cells. 
     
     
         3 . The cell line of  claim 2 , wherein the genetic modification includes inducing overexpression of a c-myc protein fused to a ligand-binding domain of a nuclear receptor. 
     
     
         4 . An assay for selecting a neurogenic agent, said assay comprising:
 (i) a stable human neural progenitor cell line of  claim 1 ;   (ii) an assay plate pre-coated with extracellular matrix proteins, wherein the cells of said cell line are cultured in a serum-free, mitogen free medium;   (iii) a test agent; and   (iv) a means for measuring a quantity or a proportion of neurons in the culture.   
     
     
         5 . The assay of  claim 4 , wherein the cells are plated at a density, said density being between about 2,000 and about 125,000 cells per well in a 96-well plate. 
     
     
         6 . The assay of  claim 4  wherein the test agent is selected from the group consisting of a fused imidazole; an aminopyrimidine; a nicotinamide; an aminomethyl phenoxypiperidine; and an aryloxypiperidine.