I am isolating primary fibroblasts from breast cancer tissue. they grow fine in TC flask and DMEM containing 10% FBS. for the assays, I have to move them to 24well plates and RPMI 10%FBS. afterwards, I have a drug treatment and mitomycin inactivation. after 2 days in this media the cells in the center start floating and die. but in the outer edge cells are fine. I have done this experiment several times and i haven't changed my protocols. what seems to be the problem?
Breast cancer cells produce hyaluronan. This may prevent adequate nourishment to the central cells surrounded by this glycan. This is just an idea. Try to detect hyaluronan in the media.
Tomas Koltai
Dear Tomas, this cell death is after isolation. they grow fine as monolayer in T25 and T75 flasks.
Dear Researcher
Check the IC50 value as reported earlier. If you get similar IC50 as reported. You have no problem in experimentation.I have also observed similar type of problem. In general under the stress condition cells migrate and some of cells become round and some of cell become ruptured. This showed that your studied drug worked.
Thank you
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