Human mesencephalon cell lines and methods of use therefor

Abstract

Conditionally-immortalized human mesencephalon cell lines are provided. Such cell lines, which may be clonal, may be used to generate neurons, including dopaminergic neurons. The cell lines and/or differentiated cells may be used for the development of therapeutic agents to prevent and treat a variety of neurological diseases such as Parkinson's disease. The cell lines and/or differentiated cells may also be used in assays and for the general study of mesencephalon cell development and differentiation.

1 . A method for producing a conditionally-immortalized human mesencephalon neural precursor cell, comprising: 
 (a) transfecting human mesencephalon cells plated on a first surface and in a first growth medium that permit proliferation with DNA encoding a selectable marker and an externally regulatable growth-promoting gene; and    (b) selecting the transfected cells on a second surface and in a second growth medium that permit attachment and proliferation, and therefrom producing a conditionally-immortalized human mesencephalon cell.    
     
     
         2 - 8 . (canceled)  
     
     
         9 . A method for producing neurons, comprising culturing a cell produced according to  claim 1  under conditions inhibiting expression of the growth-promoting gene.  
     
     
         10 - 17 . (canceled)  
     
     
         18 . A method for screening for an agent that modulates an activity of a protein produced by a human mesencephalon cell, comprising: 
 (a) contacting a cell produced according to the method of  claim 1  or  claim 9  with a candidate agent; and    (b) subsequently measuring the ability of the candidate agent to modulate an activity of a protein produced by the cell.    
     
     
         19 . A method for detecting the presence or absence of a protein in a sample, comprising: 
 (a) contacting a sample with a cell produced according to the method of  claim 1  or  claim 9;  and    (b) subsequently detecting a response in the cell, and therefrom detecting the presence of a protein in the sample.    
     
     
         20 . (canceled)  
     
     
         21 . A method for screening for an agent that affects human mesencephalon cell death, comprising: 
 (a) contacting a cell produced according to the method of  claim 1  or  claim 9  with a candidate agent under conditions that, in the absence of the candidate agent, result in death of the cell; and    (b) subsequently measuring the ability of the candidate agent to affect the death of the cell.    
     
     
         22 . A method for screening for a protein that regulates human mesencephalon cell death, comprising: 
 (a) altering the level of expression of a protein within a cell produced according to  claim 1  or  claim 9;  and    (b) subsequently measuring the effect of the alteration on the death of the cell, and therefrom identifying a protein that regulates human mesencephalon neural precursor cell death.    
     
     
         23 . (canceled)  
     
     
         24 . (canceled)