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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.
Sulforhodamine B is often used as a membrane-impermeable polar tracer [3] or used for cell density determination via determination of cellular proteins (cytotoxicity assay). [ 4 ] References
The Neutral Red Cytotoxicity Assay was first developed by Ellen Borenfreund in 1984. In the Neutral Red Assay live cells incorporate neutral red into their lysosomes. As cells begin to die, their ability to incorporate neutral red diminishes. Thus, loss of neutral red uptake corresponds to loss of cell viability. [3]
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]
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.
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 ...
Resazurin based assays show excellent correlation to reference viability assays such as formazan-based assays (MTT/XTT) and tritiated thymidine based techniques. [ 9 ] [ 10 ] The low toxicity makes it suitable for longer studies, and it has been applied for animal cells, bacteria, and fungi [ 10 ] for cell culture assays such as cell counting ...
An assay (analysis) is never an isolated process, as it must be accompanied with pre- and post-analytic procedures. Both the communication order (the request to perform an assay plus related information) and the handling of the specimen itself (the collecting, documenting, transporting, and processing done before beginning the assay) are pre-analytic steps.