Dear Scientists of ResearchGate,
With your help and guidance, I was able to successfully grow L929 cells and use them for reliable cytotoxicity evaluation per USP .
Now, I am updating the method to run as an XTT assay on a microplate reader. Does anyone have experience with that? The XTT assay kit is relatively easy to use and I'm in the process of determining the optimal cell density for each well in the microplate, but I can't seem to find much into on using the XTT assay for cell viability when adding a potentially cytotoxic compound.
Any feedback would be greatly appreciated. Thank you and best,
-Tim S
The XTT assay can not measure cytotoxicity but rather measures cell proliferation or a lack of cell proliferation. XTT/MTT assays are used for the measurement of cell proliferation in response to growth factors, cytokines and nutrients and can also used to measure the activity of growth inhibiting agents such as inhibitory antibodies i.e. a measure of cytostatsis. As such it does not provide an analysis of the percent or frequency of viable cells such as obtained with a trypan blue or PI analysis Nor does it provide an analysis of cytotoxicity such as would be measured with a 51Cr release assay.
You can measure a decrease in proliferation (cytostatsis) with this assay but it would be incorrect to label this a loss of viability or cytotoxicity. Hope this helps provide clarity.
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