Cultures of human CNS neural stem cells

Abstract

The invention provides a cell culture including proliferating human neural stem cells with a doubling rate faster than thirty days. The invention also provides a cell culture media for proliferating mammalian neural cells including a standard defined culture medium, a carbohydrate source, a buffer, a source of hormones, one or more growth factors that stimulate the proliferation of neural stem cells, and LIF. The invention also provides a method for protecting, repairing or replacing damaged tissue comprising transplanting mammalian neural stem cells formed into neurospheres. The invention also provides a cell culture of differentiated human neural stem cells where the cells are glioblasts. The invention also provides a method of differentiating human neural stem cells in culture media.

1 . A cell culture comprising: 
 (a) a culture medium containing one or more predetermined growth factors effective for inducing multipotent central nervous system (CNS) neural stem cell proliferation, wherein said culture medium comprises at least 0.1 ng/ml LIF; and    (b) human multipotent CNS neural stem cells wherein: 
 (i) the cells are grown in culture medium containing one or more predetermined growth factors effective for inducing multipotent CNS neural stem cell proliferation;  
 (ii) the population comprises cells which stain positive for nestin;  
 (iii) in the presence of differentiation-inducing conditions, the cells produce progeny cells that differentiate into neurons, astrocytes, or oligodendrocytes; and  
 (iv) the cells have a doubling rate faster than 30 days.  
   
   
   
       2 . The cell culture of  claim 1 , wherein the cells have a doubling rate of 5-10 days.  
   
   
       3 . A method for proliferating human multipotent central nervous system (CNS) neural stem cells comprising: 
 proliferating multipotent CNS human neural stem cells in serum-free culture medium containing one or more predetermined growth factors effective for inducing multipotent CNS neural stem cell proliferation and at least 0.1 ng/ml LIF, wherein:    (a) the population comprises cells which stain positive for nestin; and    (b) in the presence of differentiation-inducing conditions, the cells produce progeny cells that differentiate into neurons, astrocytes, and oligodendrocytes.    
   
   
       4 . The method of  claim 3 , further comprising the step of: exposing the proliferated cells of  claim 3  to differentiation-inducing conditions or to an appropriate tissue environment, to produce progeny cells that differentiate into neurons, astrocytes, and oligodendrocytes.  
   
   
       5 . The method of  claim 3 , wherein the cells have a doubling rate faster than 30 days.  
   
   
       6 . The method of  claim 3 , wherein the culture medium contains at least 10 ng/ml LIF.  
   
   
       7 . The method of  claim 5 , wherein the culture medium contains at least 10 ng/ml LIF.