Noncolorimetric histoculture method for predicting drug response of tumors
AssigneeAntiCancer, Inc.
InventorsKenneth M. Connors, Hui-Yan Guo
Life of this patent
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Abstract
A method of evaluating the effectiveness of drugs in inhibiting the growth of tumor cells. A sample of a tumor is histocultured, a drug to be evaluated is added to the sample and the sample is incubated. A suitable tetrazolium salt is added and a frozen section of the sample is prepared. The section is stained with a fluorescent dye. The section is exposed to polarized light and the reflected light is measured. Then the section is exposed to fluorescent light and the light emitted by the dye is measured. These measurements are compared to a control and/or measurements from tests using other drugs and the relative effectiveness of the drug is evaluated, preferably by pixel analysis.
Claims24
A method of evaluating the effectiveness of drugs in inhibiting the growth of tumor cells which comprises the steps of:
- providing a sample of a tumor;
- histoculturing said sample on a support matrix;
- adding a selected quantity of a drug to be evaluated to said histocultured sample;
- histoculturing said sample with said added drug;
- removing said added drug from said histocultured sample;
- adding a selected quantity of a tetrazolium salt to said histocultured sample;
- histoculturing said sample with said added tetrazolium salt;
- removing said added tetrazolium salt from said histocultured sample;
- preparing a frozen section of said sample;
- staining said section with a fluorescent dye;
- exposing said section to polarized light and measuring a first light reflected by said section which is proportional to the number of living cells in the sample;
- exposing said section to light of wavelengths to which said dye responds and measuring a second fluorescent light emitted by said section which is proportional to the total number of cells present in the sample;
- and
- comparing the relative amounts of said first and second light measurements;
- whereby the effectiveness of said drug tested is determined by calculating the ratio of said first light to said second light and comparing said ratio to a control value to obtain the amount of drug-induced inhibition.
A method of measuring the relative effectiveness of selected drugs in inhibiting growth of tumor cells which comprises the steps of:
- providing first and second samples of histocultured tumors;
- adding a selected amount of a drug to said first sample;
- histoculturing said first and second samples;
- removing said added drug from said first sample;
- adding a selected quantity of a tetrazolium salt to said first and second samples;
- histoculturing said samples with said added tetrazolium salt;
- removing said added tetrazolium salt from said samples;
- preparing a frozen section of each sample;
- staining each section with a fluorescent dye;
- exposing each section to polarized light and measuring the polarized light reflected by each sample;
- exposing each section to light of wavelengths to which said dye responds and measuring the fluorescent light emitted by each of said samples;
- and
- comparing the results of said light measurements;
- whereby the relative effectiveness of said drug is determined by calculating a first ratio of said first light to said second light of said first sample and a second ratio of said first light to said second light of said second sample and comparing said first and second ratios.
Dependencies are inferred from the claim text.