Dear all:
Recently, I tried to induce alcohol toxicity to FL83B cell, and I followed the literature which indicates that LD50 of EtOH is 5.5%(v/v) to FL83B cells. Last week I used CCK-8 to determine their survival rates in series EtOH-contained medium (after 6h-treating), and I got no difference between all groups; therefore, I checked those cells with microscope and got the picture as bellow. There are no any damage to those cells even up to 1000mM (about 6%). I'm wondering why? Could any one give me some ideas? Any experience on it?
Crying for help
appreciate for your kindness!
hi, perhaps your EtOH evaporates from media as it is much mre voletile than water? Maybe you shoud use water with EtOH to humidify your incubator. I really have no other idea, just a shot in the dark...
Hi Y-S S,
If your cells have usual metabolization capacities, I agree with Radoslaw that the observed cell survival should be caused by evaporation of acool which could be quite important at 37°C for 6 hours...
I suggest that you evaluate alcool concentration in a simulation of your experiment, but just in distilled water with densitometer determination of alcool content. And then maybe add some alcool from time to time in your real experiment to maintain correct concentration.
Just a suggestion
Good Luck
Patrick
Thanks for all suggestion you gave, I will check my concentration again. Maybe I should repeat the exp. more times.
Thank you again.
hi,
Normally cell lines are not resist higher conecentration of ethanol than 2% aprox.
As others researchers say you EtOH evaporates from culture media. This is the possiblity than I think ...
Good luck, Joan.
Some cells are remarkably resistant to ethanol. Some normal hematopoietic progenitor cells, for example, show LD50s of 4-5% with continuous exposure over several days (see Meagher et al, New England Journal of Medicine, 307: 845-849, 1982 and Levine et al, British Journal of Haematology 62: 345-354, 1986).
It is also possible that the CCK-8 assay, a short-term dye exclusion assay is less sensitive than the colony assays used by other investigators. In other words, if you assay your ethanol-treated cells by in vitro clonal assay,you might well see some evidence of toxicity.
- Badges
- Science topic