Methods of identifying cytotoxic effects in quiescent cells
OwnerBruce Littlefield, Kathleen Salvato +1inventors
InventorsBruce Littlefield, Kathleen Salvato, Laura Rudolph-Owen
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Abstract
The present invention provides a system for measuring the cytotoxicity of a test compound against quiescent cells. The inventive methods and reagents may also be used to determine whether a compound, such as a chemotherapeutic agent, will have undesired toxicity toward normal tissues, which generally do not include a significant number of cells that are proliferating. Thus, the assay of the invention can be used to identify in vitro compounds that demonstrate a good therapeutic index, thus, accelerating the development of drugs that are clinically useful.
Claims69
A method of determining the cytotoxicity of a test compound to quiescent cells, comprising the steps of:
- a) growing plated cells to confluence in a tissue culture medium containing between about 5% and about 20% animal sera;
- b) washing the cells one or more times;
- c) serum starving the cells for at least about six hours in a tissue culture medium that contains between about 0% and about 0.5% animal sera, thereby obtaining a quiescent cell culture;
- d) incubating the quiescent cell culture with the test compound for at least about six hours;
- and
- e) determining the viability of the cells, thereby determining the cytotoxicity of the test compound.
A kit for measuring the cytotoxicity of a test compound towards quiescent cells, comprising:
- a) a compound that inhibits cell proliferation;
- and
- b) a cytotoxic compound.
Dependencies are inferred from the claim text.