Hi, I culture AGS cells human gastric cancer cells. I use materials 'flavonoid' for inhibiting cancer cells. I use DMSO for solvent, 0.5% of total volume in RPMI. Anyway, i treated with various concentrations of materials for 24 hour for protein extraction in 75t-flask. (Cells were about 80% in falsk.) I always process this course. However, Cells were disappeared after just one day in treated flask recent experiment. All cells were floating medium. (Cells were just 10-20% in falsk, what was worse, it was Control.) Have you ever been experienced this situation? I don't know why this happened.
Were all of the flasks this way or just one flask (control)? If just one, this is usually due to a random single event contamination of some kind. If it is all of the flasks then you may have a contaminated stock solution/materials.
As suggested above Eric L Thompson, this may be the possibility. Another possibility is that your cells are over confluent. As you treated cells, all cells may be detached.
Eric L Thompson, All of the flask were disappeard. But i made new stock solution, and used that..So i think materials were possible contaminated.. If that was ture, i would check Ventilation license..
Ravi P. Cholia, I cultured cells in 60 dish. Then, there were just floating, didn't adhere floor. That was possoble, because over confluent too?
DId your control medium also contain 0.5 % DMSO? The DMSO concentration is critical already at 0.1%.
Angela M Otto, is taht true? i was processed all of my experiment '0.5 % of DMSO'.... i read both 0.1% and 0.5% DMSO in medium... so i selected that 0.5% DMSO.
This is not only my experience. May I refer you to a previous discussion: https://www.researchgate.net/post/Does_anybody_know_what_is_the_safe_solution_of_DMSO_for_cell_cultures
Also, DMSO is cell membrane permeable and could affect adhesion properties. I suggest you check studies using similar compounds as those that you're testing for appropriate solvents (concentration).
Good success!
I cannot say about gastric cancer cells (never work with them), but in case of prostate or breast cancer cells you should split them at 75-80% of confluence. In normal cells contact inhibition usually leads to quiescence, but cancer cells will die. It is better to start the experiment no more then 50% of confluence (sometimes even earlier, it depends how fast cells are growing and the time points of experiment).
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