Method for using division arrested cells in screening assays
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 - 24 . (canceled)
25 . A method for determining if a substance of interest has an effect on a type of cell, comprising:
(i) contacting said substance of interest to a sample of said type of cell, wherein cells in said sample have had their division arrested, so the either their mitotic or meiotic cycle has been stopped, and cellular division can no longer take place,
(ii) comparing any change in said sample of (i) to a sample of cells which have had their division arrested in the same way but have not been contacted with said substance of interest, wherein a difference between said samples is indicative of an effect of said test substance on said type of cell.
26 . The method of claim 25 , wherein said samples of cells in (i) and (ii) have been cryopreserved.
27 . The method of claim 25 , wherein said type of cell is a eukaryotic cell.
28 . The method of claim 25 , wherein said type of cell is a prokaryotic cell.
29 . The method of claim 25 , wherein division of said type of cell has been arrested during mitosis.
30 . The method of claim 29 , comprising arresting mitosis by treatment with mitomycin C.
31 . The method of claim 29 , comprising arresting mitosis by treatment with gamma irradiation.
32 . The method of claim 25 , wherein division of said type of cell has been arrested during meiosis.
33 . The method of claim 25 , wherein said substance of interest is a protein and said cell has been transformed or transfected with a nucleic acid molecule which expresses a protein with which said test substance interacts.
34 . The method of claim 33 , wherein said nucleic acid molecule comprises a recombinant virus vector.
35 . The method of claim 33 , wherein said cell has been transformed or transfected with RNA.
36 . The method of claim 25 , wherein said cell has been transduced with a protein, peptide, or protein containing molecule.
37 . The method of claim 25 , wherein said cell is an NIH3T3 cell.
38 . The method of claim 25 , wherein said method comprises measuring induction of Ca2+ immobilization.
38 . The method of claim 25 , wherein said method comprising measuring induction of Ca 2+ immobilization.
39 . The method of claim 25 , wherein said cell is an HEK293 cell.
40 . The method of claim 25 , wherein said cell has been transformed or transfected with a nucleic acid molecule which encodes a reporter molecule, wherein said reporter molecule gives a detectable signal upon modulation of said test protein with said type of cell.
41 . The method of claim 40 , wherein said reporter molecule is expressed due to activation of a molecule expressed in said cell, whose activation is caused by interaction of said substance of interest and a molecule expressed by said type of cell.
42 . The method of claim 41 , wherein said reporter molecule is luciferase, or green fluorescent protein.
43 . The method of claim 25 , said method comprising a competitive assay wherein said substance of interest is admixed with a compound known to effect a molecule expressed by said type of cell, and determining any change in the known effect as a determination that said substance of interest effects said type of cell.