I want to check the effect of chemotherapeutic dug on breast cancer cell line, which possible tests can I perform to check either cells are showing resistant to the drug or not?
Thank you
The primary thing to do is to check for proliferation curve. Compare the cell proliferation in vehicle control vs drug at various concentration to find out IC50. Once you establish that, plan for next set of experiments. Hope this helps.
Following Gaurang's idea you can check for cell viability (or cytotoxicity) with different concentrations and compare them with normal cells. If both IC50 are close each to other, that would mean that your drug is equally toxic for normal and cancer cells. Therefore, it would not be very useful. If there is a substantial difference between both IC50 then you have an interesting drug that deserves further research.
Thank you so much Gaurang L Trivedi and Tomas Koltai for your precious suggestions. I will count cell viability after providing them various concentrations of different drugs (selected). My supervisor advised me MTT assay for this. So I want to know is there any other test/assay which can I perform?
Thank you.
We also use flow cytometry based assays with propidium iodide and flow cytometry count standards.
Gaurang L Trivedi Can you please share link of the protocol.
Thank you
Look what I shared: Cell Viability Reagents for Flow Cytometry, Microscopy & ELISA | Bio-Rad @MIUI| https://www.bio-rad-antibodies.com/cell-health-cell-viability-dyes-reagents.html
Try this link
Shumaila you can perform live cell imaging with drug concentrations, if IncuCyte is available in the lab.
Colony forming assay can also be performed just like MTT assay (using 6 well plate and count colonies over a period of couple of weeks).
Always remember to use controls.
you can also go western blotting for DNA damage prediction using different markers (antibodies).
There are a lot of things to do once you get handy with cell tissue culture experiments.
Good Luck
BIOPSY - MRI SCAN / CANCER MARKER IN BLOOD after applying Chemothepeutic Agents .
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