Method for using division arrested cells in screening assays

Abstract

Division arrested cells are used in screening assays to determine the effect of a substance of interest on the cells. The division arrested cells can be used in drug screening assays, signal transduction assays, and are especially useful in large scale, high throughput assays.

1 . A method for determining effect of a substance of interest on a type of cell, comprising contacting the substance of interest to a sample of said type of cell, wherein the cells in said sample have had their division arrested, and determining a change in said sample of said cells following contact with said substance as a determination that said substance has an effect on said type of cell.  
     
     
         2 . The method of  claim 1 , wherein said cell is a eukaryotic cell.  
     
     
         3 . The method of  claim 1 , wherein said cell is a prokaryotic cell.  
     
     
         4 . The method of  claim 1 , wherein division of said cell has been arrested during mitosis.  
     
     
         5 . The method of  claim 1  wherein division of said cell has been arrested during meiosis.  
     
     
         6 . The method of  claim 1 , wherein said cell has been transformed or transfected with a nucleic acid molecule which expresses a protein of interest.  
     
     
         7 . The method of  claim 6 , wherein said cell has been transformed or transfected with a recombinant virus.  
     
     
         8 . The method of  claim 1 , wherein said cell has been transformed or transfected with RNA.  
     
     
         9 . The method of  claim 1 , wherein said cell has been transformed or transfected with an antisense nucleic acid molecule.  
     
     
         10 . The method of  claim 1 , wherein said cell has been transduced with a protein, peptide, or protein containing molecule.  
     
     
         11 . The method of  claim 1 , wherein said cell is an NIH3T3 cell.  
     
     
         12 . The method of  claim 1 , said method comprising measuring induction of Ca 2+  immobilization.  
     
     
         13 . The method of  claim 1 , wherein said cell has been transformed or transfected to produce 5HT2c.  
     
     
         14 . The method of  claim 1 , wherein said cell expresses a G-protein coupled receptor.  
     
     
         15 . The method of  claim 1 , wherein said cells express ion channels.  
     
     
         16 . The method of  claim 14 , wherein said G-protein coupled receptor is muscaranic acetylcholine receptor.  
     
     
         17 . The method of  claim 1 , wherein said cell is an HEK293 cell.  
     
     
         18 . The method of  claim 1 , wherein said cell expresses a .beta.2 adrenergic receptor.  
     
     
         19 . The method of  claim 1 , wherein said cell has been transformed or transfected with a nucleic acid molecule which encodes a reporter molecule.  
     
     
         20 . The method of  claim 19 , wherein said nucleic acid molecule is activated via activation of a molecule expressed in said cell.  
     
     
         21 . The method of  claim 19 , wherein said reporter molecule is luciferase, or green fluorescent protein.  
     
     
         22 . The method of  claim 1 , said method comprising a competitive assay wherein said substance of interest is admixed with a compound known to affect to said cell, and determining any change in the known affect as a determination that said substance of interest affects said cell.  
     
     
         23 . A kit useful in screening a substance of interest to determine its affect on a cell type of interest, comprising a container means which holds a separate portion of each of: (i) a substance which arrests cell division of said cell; and, (ii) a substance known to interact with said cell and to provoke a known affect.  
     
     
         24 . The kit of  claim 23 , further comprising a separate portion of (iii) a substance which affects a function other than cell division in said cell.