Established cell line of microglia

Abstract

A subcultivatable, established microglia having the following properties. (a) Form: Both or either of a macrophage-like or spherical form in the presence of a granulocyte-macrophage colony stimulation factor and a branched form similar to branched microglia present in the brain in the absence of the factor. (b) Functional characteristics: specific affinity for the brain highly poor phagocytic action. (c) Cell proliferation: proliferative depending upon a granulocyte-macrophage colony stimulation factor. Preparation, separation, and screening methods of the microglia, a pharmaceutical composition using the microglia, and a method for treatment of cerebral diseases using the composition.

What is claimed is:  
     
         1 . A subcultivatable, established cell line of microglia.  
     
     
         2 . The established cell line of microglia according to  claim 1 , which has the following properties: 
 (a) form: having either a macrophage-like or globular form in the presence of granulocyte-macrophage colony-stimulating factor, or in the absence of said factor, a branched form similar to branched microglia present in the brain, or both of the above forms;    (b) functional characteristics: having specific affinity for the brain, and having a strong phagocytic ability; and    (c) cell growth ability: growing depending on granulocyte-macrophage colony-stimulating factor.    
     
     
         3 . A method of separating the established cell line of microglia described in  claim 1  or  2  from microglia in the presence of a cytokine.  
     
     
         4 . The method according to  claim 3 , wherein the cytokine is GM-CSF.  
     
     
         5 . The method according to  claim 4 , wherein GM-CSF is a genetic recombinant one.  
     
     
         6 . The method according to any one of  claims 3  to  5 , which is carried out in the presence of IL-3 and/or a culture supernatant of purified astrocytes.  
     
     
         7 . A pharmaceutical carrier comprising the established cell line of microglia described in  claim 1  or  2 .  
     
     
         8 . The established cell line of microglia according to  claim 1  or  2 , comprising a gene or a drug introduced into it.  
     
     
         9 . The established cell line of microglia according to  claim 8 , comprising a gene introduced into it.  
     
     
         10 . A pharmaceutical composition comprising the established cell line of microglia of  claim 8  or  9  and a pharmaceutical carrier.  
     
     
         11 . The pharmaceutical composition according to  claim 10 , which is an agent for treatment of cerebral diseases.  
     
     
         12 . A method of screening a microglia having a gene introduced into it, comprising introduction of an extraneous gene and a fluorescent protein-expressing gene into a microglia and subsequent screening, by the fluorescent protein of the microglia having the gene introduced into it.  
     
     
         13 . The method according to  claim 12 , wherein the fluorescent protein-expressing gene is derived from a jellyfish.  
     
     
         14 . A process for producing a microglia having a gene introduced into it, comprising introduction of an extraneous gene and a fluorescent protein-expressing gene into a microglia and subsequent screening, by the fluorescent protein, of the microglia having the gene introduced into it.  
     
     
         15 . The method according to  claim 13  or  14 , wherein the fluorescent protein-expressing gene is derived from a jellyfish.  
     
     
         16 . A method of treating cerebral diseases, which comprises using the pharmaceutical composition of  claim 10  or  11  to deliver a drug or gene specifically to the brain.