Cell culturing methods and apparatus
AssigneeBrown University Research Foundation
InventorM. Boris Rotman
Life of this patent
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Abstract
A system and protocol for performing cytotoxicity studies including apparatus and methods for culturing the biopsied cells, for providing oxygenated nutrients, for removing cell-waste products, for introducing a fluorogenic substrate, for introducing cytotoxic agents including anticancer drugs, for measuring the released fluorescence and for measuring intracellular accumulation of fluorescein. Cytotoxicity can be determined by measuring fluorescence in the efflux of the vessel and concommitantly by direct photometric comparisons of the cells in the vessel before and after exposure to the cytotoxic agent.
Claims14
A cytotoxicity assay system comprising a cell culturing vessel having a cell enclosure into which cells can be introduced and cultured;
- nutrient-transporting means for transporting nutrients to the cells, the nutrient-transporting means include a source of nutrients and at least one fluid permeable membrane, connecting said source of nutrients to said cells, across which nutrients can migrate to nourish the cells within the vessel;
- fluorogenic substrate-transporting means for transporting a fluorogenic substrate to the cell compartment, so that living cells within the vessel will take up said substrate and accumulate a characteristic fluorescence;
- means for introducing a therapeutic agent into the cell compartment;
- and
- means for measuring cytotoxicity within the cell compartment, including means for detected changes in the fluorescence of the cells within the vessel.
A method of assaying the sensitivity of biopsied cells to therapeutic agents, the method comprising:
- a. culturing the cells in a vessel;
- b. contacting the cells with a fluorogenic substrate whereby living cells accumulate a characteristic amount of fluorescence;
- c. introducing the agent into the vessel;
- and
- d. measuring changes in the fluorescence by photometrically analyzing the intensity of fluorescence within the vessel as an indicator of the sensitivity of the cells to the agent.
Dependencies are inferred from the claim text.