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 method for implanting neural stem cells in the central nervous system of a patient comprising transplanting a therapeutically effective amount of human neural stem cell cultures comprising human neural stem cells that have a doubling rate faster than 30 days, wherein the cultures comprise cells which stain positive for nestin, and wherein in the presence of differentiation-inducing conditions said cells produce progeny cells that differentiate into neurons, astrocytes, or oligodendrocytes.  
     
     
         2 . The method of  claim 1 , wherein the human neural stem cells have a doubling rate of 5-10 days.  
     
     
         3 . The method of  claim 1 , wherein the human neural stem cells are substantially formed into neurospheres of a diameter between 10-500 μm.  
     
     
         4 . The method of  claim 1 , wherein the cultures comprise between 10,000 and 1,000,000 cells/μl.  
     
     
         5 . The method of  claim 1 , wherein the cells are proliferated in suspension culture.  
     
     
         6 . The method of  claim 1 , wherein the cells are proliferated in an adherent culture.  
     
     
         7 . The method of  claim 1 , wherein the progeny of said neural stem cells are genetically modified.  
     
     
         8 . A method for implanting neural stem cells in the central nervous system of a patient comprising: 
 a) obtaining a culture comprising human neural stem cells having a doubling rate faster than 30 days, wherein the cultures comprise cells which stain positive for nestin, and wherein in the presence of differentiation-inducing conditions said cells produce progeny cells that differentiate into neurons, astrocytes, or oligodendrocytes;    b) exposing said culture to differentiation inducing-conditions; and    c) transplanting a therapeutically effective amount of said neurons, astrocytes, or oligodendrocytes to the central nervous system of the patient.