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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.
A microtiter plate after an MTT assay. Increasing amounts of cells resulted in increased purple colouring. The MTT assay is a colorimetric assay for assessing cell metabolic activity. [1] [2] NAD(P)H-dependent cellular oxidoreductase enzymes may, under defined conditions, reflect the number of viable cells present.
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 ...
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 viability assay is an assay that is created to determine the ability of organs, cells or tissues to maintain or recover a state of survival. [2] Viability can be distinguished from the all-or-nothing states of life and death by the use of a quantifiable index that ranges between the integers of 0 and 1 or, if more easily understood, the range ...
A cell rounding assay (cytotoxicity assay) has been developed to diagnose C. difficile infection. [11] Enzyme-linked immunosorbent assays (ELISAs) have been used to detect TcdA and TcdB with specific antibodies. When used with an ELISA, the cytotoxicity assay is the "gold standard" when used on Vero cells for C. difficile diagnosis. [11]
[2] [3] The cell culture assay is regarded as a "gold standard" for detecting toxicity in C. difficile because a small quantity of toxin B is capable of causing cell rounding (Fig. 4), thus, it is a major advantage of clinical laboratories to make correlations with the CDAD caused by TcdB.