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A cell culture assay is any method used to assess the cytotoxicity of a material. [1] [2] This refers to the in vitro assessment of a material to determine whether it releases toxic chemicals in the cell. It also determines if the quantity is sufficient to kill cells, either directly or indirectly, through the inhibition of cell metabolic pathways.
Cytotoxicity can also be monitored using 3-(4, 5-Dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide or with 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), which yields a water-soluble product, or the MTS assay. This assay measures the reducing potential of the cell using a colorimetric reaction.
Cytotoxicity can be quantified by measuring the amount of label in solution compared to the amount of label that remains within healthy, intact cells. The classical method of detecting this is the chromium-51 [51 Cr] release assay; the sulfur-35 [35 S] release assay is a little used radioisotope-based alternative. Target cell lysis is ...
A high throughput assay can be either an endpoint or a kinetic assay usually done on an automated platform in 96-, 384- or 1536-well microplate formats (High Throughput Screening). Such assays are able to test large number of compounds or analytes or make functional biological readouts in response to a stimuli and/or compounds being tested. [6]
Complement-dependent cytotoxicity (CDC) is an effector function of IgG and IgM antibodies.When they are bound to surface antigen on target cell (e.g. bacterial or viral infected cell), the classical complement pathway is triggered by bonding protein C1q to these antibodies, resulting in formation of a membrane attack complex (MAC) and target cell lysis.
A protocol for the MAT test, using cultured cells, is described in the European Pharmacopoeia. [16] A recent study employing genetically engineered monocytes was able to significantly enhance the sensitivity of monocyte-based detection assays by bringing down the assay-completion time from more than 20 hours to 2–3 hours. [17]
In phase one, effector CTLs are generated from CTL precursors. The CTL precursors include naive T c cells since they are incapable of killing target cells. After a precursor cell has been activated, it can then differentiate into a functional CTL with cytotoxic activity.
In vitro toxicity testing is the scientific analysis of the toxic effects of chemical substances on cultured bacteria or mammalian cells. [1] In vitro (literally 'in glass') testing methods are employed primarily to identify potentially hazardous chemicals and/or to confirm the lack of certain toxic properties in the early stages of the development of potentially useful new substances such as ...