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 . A method for determining if a drug candidate has an effect on a mammalian cell, the method comprising:
(a) contacting the mammalian cell with the drug candidate, and
(b) determining whether the drug candidate has an effect on the mammalian cell by measuring a change in the mammalian cell,
wherein either the meiotic or mitotic cycle of the mammalian cell has been stopped and cellular division can no longer take place.
2 . The method of claim 1 , wherein activation of a G-protein coupled receptor is measured.
3 . The method of claim 2 , wherein the G-protein coupled receptor is overexpressed by the mammalian cell.
4 . The method of claim 2 , wherein the G-protein coupled receptor is selected from the group consisting of:
(a) a serotonin receptor;
(b) a muscaranic acetylcholine receptor; and
(c) a β2 adrenergic receptor.
5 . The method of claim 1 , wherein the change measured in the mammalian cell results from Ca 2+ mobilization.
6 . The method of claim 1 , wherein the change measured in the mammalian cell is determined by measuring the activity of a reporter.
7 . The method of claim 6 , wherein the reporter is selected from the group consisting of secreted alkaline phosphatase, luciferase, and green fluorescent protein.
8 . The method of claim 1 , wherein the determination whether the drug candidate has an effect on the mammalian cell is made by comparison of the mammalian cell before and after the mammalian cell are contacted with the drug candidate.
9 . The method of claim 1 , wherein the mammalian cell is a cell selected from the group consisting of:
(a) a Chinese hamster ovary cell;
(b) a NIH3T3 cell;
(c) a HEK293 cell; and
(d) a M1WT3 cell.
10 . The method of claim 1 , wherein the meiotic or mitotic cycle of the mammalian cell has been stopped by irradiation of the mammalian cell or exposure of the mammalian cell to mitomycin C.