cytotoxicity Protocols

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

Analysis of DNA Fragmentation Using the JAM Assay. By Shailaja Kasibhatla et al., The JAM assay is based on labeling nuclear DNA of cycling cells with [3H]thymidine and harvesting samples on glass fiber filters. Apoptosis will generate DNA fragments small enough to pass through the glass fiber filter, resulting in decreased radioactivity of the particular sample. Cell-mediated cytotoxicity or cell killing mediated by cytotoxic T lymphocytes (CTL) can also be measured by this technique. - [Read Analysis Of DNA Fragmentation Using The JAM Assay (Subscription Required)]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

Bovine spermatozoa cytotoxicity test. - [Read Bovine Spermatozoa Cytotoxicity Test]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Choosing a cell viability or cytotoxicity assay from among the many different options available can be a challenging task. Includes information on: Establishing an In Vitro Model System; Choosing an Endpoint to Measure; Characterizing Assay Responsiveness; Determining Dose and Duration of Exposure; Homogeneous Assays for Multiwell Formats and Automated Screening; Additional Factors to Consider When Choosing a Cell Viability Assay; Cell Viability Assays that Measure ATP Protocol; etc.. - [Read Cell Viability Information For Protocols and Applications]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The potential cytotoxicity of compounds under hypoxic conditions is determined by exposing cell cultures to test compounds in a low oxygen atmosphere.  Subsequent cell survival is determined by the MTT and methylene blue colorimetric assays. - [Read Colorimetric Cytotoxcity Assays for Anchorage Dependent Cells Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

This bioassay utilizes cultured Hepa-lclc7 (Hepa-1) mouse hepatoma cells to assess the CYPlA1-inducing potency or cytotoxicity of pure test chemicals or environmental samples.  In the Hepa-l induction test , the CYPlA1-inducing potency of the test sample is detected as increased aryl hydrocarbon hydroxylase (AHH) and 7-ethoxyresorufin O-deethylase (EROD) activities. - [Read CYP1A1-Inducing Potency and Cytotoxicity Test in the HEPA-1 Mouse Hepatoma Cell Line]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

Cell-based assays are important tools for contemporary biology and drug discovery because of their predictive potential for in vivo applications.This assay gives ratiometric, inversely proportional values of viability and cytotoxicity (Figure 4.15) that are useful for normalizing data to cell number. Also, this reagent is compatible with additional fluorescent and luminescent chemistries. - [Read Determining Number of Live and Dead Cells in Cell Population: Cytotoxicity Assay Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

This method measures the leakage of DNA and lactate dehydrogenase 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). - [Read Human Lymphocyte Cytotoxicity Assay Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

Skin fibroblasts are incorporated into 3-D collagen lattices containing the test compounds.  An inhibition of lattice contraction indicates a possible toxic effect which is verified by trypan blue exclusion for cell viability. - [Read Human Skin Fibroblast/Collagen Lattice Cytotoxicity Test]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

The results of cytotoxicity tests in primary cultures of rat hepatocytes and in MDBK and McCoy cells can be used to predict the in vivo 4-wk maximum tolerated dose in rats and dogs.  A correlation between in vitro cytotoxicity, as measured in this system, and LD50 values in rats and mice has also been established. - [Read In Vitro Prediction of the Maximum Tolerated Dose Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

The basis of this procedure is that two specific cell type preparations may be isolated, exposed separately to various compounds over a range of concentrations, and the cytotoxicity of these determined.  Parameters deemed indicative of a cytotoxic effect include a reduction in de novo protein synthesis and decreased glucose and fatty acid metabolism.  A cytotoxic effect may indicate that a chemical is likely to be nephrotoxic in vivo. - [Read Isolated Rat Glomeruli and Proximal Tubules]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

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. - [Read LS-L929 Cytotoxcitiy Test]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

Potential embryotoxicity is assessed by monitoring the effect of the test compound on total protein synthesis, and DNA synthesis in cultured human foetal lung fibroblasts.  Rat lung epithelial cells can be used to determine cytotoxicity of select compounds because of their ability to metabolise xenobiotics. - [Read Lung Cell Assay Protocol]

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The latest generation of Promega cell-based assays includes luminescent and fluorescent chemistries to measure markers of cell viability, cytotoxicity and apoptosis, as well as to perform reporter analysis. Using these tools researchers can investigate how cells respond to growth factors, cytokines, hormones, mitogens, radiation, effectors, compound libraries and other signaling molecules. The protocols provided are guidelines for multiplexing cell-based assays & are intended as starting points. - [Read Multiplexing Cell Viability Assays Protocols]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

The cytotoxic effect of chemicals upon mammalian cells, such as BALB/c 3T3 and HepG2, in culture is measured by highest tolerated dose (HTD), cell viability (Neutral Red) and total cell protein (coomassie blue). - [Read Neutral Red Cytotoxicity Assay Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

Rabbit-derived corneal cells are cultured in the presence of test compounds, the toxicity of which are determined by their effect upon cell viability.  A decrease in cell number, as measured by uptake of the dye Neutral Red, serves as an indicator of potential cytotoxicity.  This test has been proposed as a potential replacement alternative for the Draize Eye Irritation test. - [Read SIRC Cytotoxcitiy Test]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cytotoxicity Assays in Cell Culture Protocols

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). - [Read The Frame Cytotoxicity Test Kenacid Blue]