Dear all,
I think that usually wound healing assay are made in 12- or 24-well plates. However, I need to test different drugs in triplets as single treatments and some of them in combinations.
Due to 1) Some of the drugs are available in a really small quantity.
2) I need all treatments to be measured at the same time.
3) It would be even better to use less cells and less media.
So, my question is, can we do such assays in 48-well plates? And would it also be feasible if we scratch the cell monolayer with a 200 microliters yellow?
Many thanks and regards.
Hello Mark Ashraf Fahmy
Yes, wound healing assay can be performed in 48-well plate.
Yes, you may scratch the cell monolayer with 200µl pipette tip (yellow).
Some information I would like to share with you.
2. Remove the medium with mitomycin C and wash the wells once with 250µl of 1X PBS.
3. Add 250µl of 1X PBS and manually scratch the wells vertically with a 200µl yellow pipette tip. Use a new pipette tip for each well.
4. Remove the PBS and then add 250µl of the drug. Proceed further with the assay.
Best.
Dear Malcolm Nobre
Thank you for your answer, but I was planning that, we could use media supplemented with 5% only FBS instead of mitomycin C. I think this one step less
Dear Mark,
I have never used media with 5% FBS. Have you used it before in place of mitomycin C? If yes, were you satisfied with the results?
I'm not sure what you are trying to ask? Media containing 5-15% FBS is frequently used for mammalian cell/tumor cell tissue culture; and Mitomycin C is a highly cytotoxic DNA cross-linking agent, used as an anticancer agent. They serve two entirely different functions, and can't replace each other.
However, co-transfection with MCRA (the gene for mitomycin C resistance) can be used as a selection marker in tissue culture experiments. see the below reference.
Novel selection marker for mammalian cell transfection
Raymond P Baumann , David H Sherman, Alan C Sartorelli
- DOI: 10.2144/02325st03