I have to try a compound in these cell lines. I used to do MTT tests, but I wonder if x-CELLigence could be as sensitive as a MTT test?
MTT test is a colorimetric test, so it is not very sensitive compared to other tests using for example luminescence.
If you need sensitivity or you have a low amount of cells and you have a luminometer i suggest you CellTiter Glo Assay from Promega.
Dear Carolina,
cell impedance based cell analysis is a very sensitive tool for assessment of toxicity provided the involved mechanisms include the cytoskeleton. The continuous parameter recording additionally gives you an idea on the dynamic of the cellular drug response which is much more informative than an end point assay like MTT. Bionas also offers an impedance based cell analyzing assay in 96-well format (Bionas Discovery adcon reader) which is comparable to the 96-well x-CELLigence system. We treated A549 cells with staurosporin in different concentrations for 24 hours and finished with CCK-8 assay. Comparing the IC50 from the adcon reader impedance assay with CCK-8 assay we performed in parallel in TPP plate and adcon plate we got similar results.
The applicability of impedance based assays depends on involved mechanisms. If your compound effects the mitochondria it is better to monitor cellular activity of oxygen consumption. When monitoring cell impedance as the only parameter you will miss the early story of toxic manifestations. Effects on glycolysis will only be seen if you monitor the proton release activity. The Bionas Discovery 2500 system monitors these three parameters in parallel and gives you a more comprehensive picture on cellular drug response. For more details please go to www.bionas-discovery.com. or get back to me. Bionas specialises in label-free cell-based real time assays and offers both systems and services.
Thank you Fritz Evert for all the information. My compound is an Akt inhibitor,and so I would like to test it in hepatocarcinoma cell lines,as I did in leukemia cells. The difference is that the leukemia cells that we have are in suspension,while the hepatocarcinoma cells are adherent cells. X-CELLigence can be used only with adherent cells. With suspension cells I did MTT tests,with the hepato cell lines I would like to an experiment like a MTT using x-CELLigence, for example monitoring at first the proliferation of cells and then, after 24 or 48 hrs, adding the compound at different concentrations. I wonder therefore if it could be as sensitive as the MTT assay. I'll try!
Thank you for all!
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