Detailed description of the procedure to culture HEK 293 cells and the assays that are used to assess their cell viability (cell lysis and metabolic activity) after 24 h toxicant exposure.
The inhibition of CHO cell proliferation provides an overall assessment of the toxicity of the test substance.
The effect of chemicals on the activity of the plasma membrane-bound Na+/K+ -ATPase isolated from Chinese Hamster Ovary (CHO) cells is used as a measure of their toxicity.
Changes in the balance of cytoskeletal proteins after exposure to test compounds can be detected by indirect immunofluorescence microscopy and quantitative biochemical methods.
The ability of cultured cells to synthesise protein is used to assess the effect of a test compound on cellular anabolic competence.
This method measures the leakage of DNA and lactate dehydrogenase (LDH, EC. 1.1.1 27) from lymphocytes into the surrounding medium as an indicator of cytotoxicity. This method also includes an assay of intracellular (mitochondrial) diaphorase as a measure of cellular activity (MTT assay).
This simple cell culture-based cytotoxicity test (in which cell viability is determined by uptake of the dyes ethidium bromide and fluorescein acetate) has been developed as a general test for acute toxicity.
Tumour cell lines cultured as aggregates can be utilised for in vitro radiosensitivity and/or chemosensitivity tests. Chemical effects are monitored by studying the changes in spheroid diameter measured by laser diffraction.
This protocol provides a generic description of a simple assay, which can be used to determine the viability/number of cells in culture. The qunatitative measurement is made through a formation of a coloured product (in a mitochondria-dependent reaction) to which the cell membrane is impermeable.
AVEC-DIC microscopy in combination with mitochondria-specific fluorescence allows a quantitative analysis of cell organelle dynamics and fine structure in cell cultures exposed to test compounds.
Steady-state intracellular daunorubicin accumulation assay adapted to a 96-well format, for assessing the P-gp (P-glycoprotein, ABCB1) inhibition in vitro.
Steady-state intracellular mitoxantrone accumulation assay adapted to a 96-well format, for assessing the BCRP (breast cancer resistance protein, ABCG2) inhibition in vitro.
The cytotoxic effect of chemicals upon cells in culture is measured by the change in total cell protein arising from the inhibition of cell proliferation (Kenacid Blue R dye-binding method).
The cytotoxic effect of chemicals upon cells in culture is measured by cell viability (neutral red uptake) method.
Membrane permeability of perfused cell cultures, as determined by the efflux of [3H]-2-deoxy-D-glucose-6-phosphate, is used as an indicator of the cytotoxic effect of chemicals.
The activating system (human liver microsomes) is separated by a semi-permeable membrane from the target cells (human mononuclear leucocytes or red cells) in order to identify cytotoxic metabolites that are capable of diffusing away from the site of production.
The absorption of UV at 260nm in a fixed volume of solubilised cells is proportional to the cell number, and therefore can be used as a simple means of obtaining a cell count. Cell counts obtained in this way can be combined with measurements of the inhibition of DNA synthesis ([3H]-thymidine incorporation) by test compounds, to produce an index of cytotoxicity.
The cytotoxic effect of test chemicals in V79 cell culture can be determined by assessing damage to the plasma membrane as determined by a nucleic acid leakage assay.
The cytotoxic effect of chemicals upon yeast (Saccharomyces cerevisiae) cells in culture is determined by inhibition of cell proliferation, as measured by cell density.
The effect of chemicals on the activity of the plasma membrane-bound H+-ATPase, isolated from yeast (Saccharomyces cerevisiae) cells, is used as a measure of their toxicity.