Chemoresistance assay - How to measure the chemoresistance of cancer cells (Mar/03/2008 )
Hi everyone ..... I need your help to find a quick, reliable, sensitive method for the measurement of chemoresistance of cancer cells that can give reproduceable results under a wide of range of chemical concentrations. I have been searching the literature and found different types of assays like Fluorescent Microculture Cytotoxicity Assay (FMCA); Adenosine Triphosphate (ATP) Bioluminescence Assay (“cell-death” assay) What type of assay should be preferred ?? What could be other types of methods for determining chomoresistance of cells ??? We are working with prostate cancer cell lines and want to know the change in response of these cells along with selective gene expression against different types of chemicals ... so what could be the best methadology ??? Plz let me know if the description lacks something. Thanks a lot and waiting for the expert responses.
Thanks a lot friends for atleast having a look at my problem... but still i have not got the help i waiting for. Am I at the wrong forum ??? or no one has got any intrest in the cell viability tests ?? Please help me with any simple and reproduceable procedures that you have tried in your labs. Thanks
i'm using the celltiterglo reagent from promega.
it is fast, easy and accurate but maybe a little bit pricey compared to others.
you plate your cells on 96well plates, treat them with serial dilutions of your drug, wait for 48h, just pipet 100µl of reagent in each well, measure, finished.
Thanks a lot Ned. This seems a bit cheaper as compared to similar products from Roche, Sigma and Lonza. We will talk about the suitability of this assay for our experiments. Thanks again
i should also mention that you need opaque plates for this assay to prevent crosstalk between the wells. i use black plates with clear bottom which is also ok., so i can also look at the cells under the microscope.
We have 96well opaque plates that we use for real-time PCR but their bottom is not clear so will not have a chance to view under the microscope. Will they work ?? or we have to buy the plates that you mentioned ??? Thanks again Ned for the intrest.
you can't culture cells in PCR plates. a possibility would be to transfer an aliquot of the luminescent cell lysate to these plates and analyze them. but then you would need a machine that can read the luminescence in the PCR plates
there are suitable plates for example from nunc or falcon.