Hello dear scientists:
When I treated the Pc12 cell line with H2O2, after 2h the cells seemed to be dead.
I didn't observe crystals, so I didn't add DMSO and left it with mtt in the incubator.
Today I checked it (after 24h) all the wells contained formazans. Also, oD was ok too. Is it a reliable result?
In short, yes it is possible. Different cells have different susceptibilities to hydrogen peroxide. But how did you measure OD after without DMSO which solubilize the formazan crystals prior to OD read at 570nm. It is best to do a multi-well culture then half viability stain after hydrogen peroxide treatment (different incubation periods or concentrations) and half for MTT. 24 hrs may be too long and 2 hrs too short since literature on pc12 shows 4-6 hrs to be ideal, i would do 4, 6, 12hrs.
Rabeah Al-Temaimi after seeing formazan I changed the media and added 5o microliter DMSO and read the plate at 570 nm. Thank you I will try your suggestion. Today I observed the same for my synergic test, at first cells, dead, but after 24h, Fromazans appeared.
literature examples (Article Neuroprotective effect of paeoniflorin on H2O2-induced apopt...
) and (Article Differential protection of pre-, co- and post-treatment of c...
) where 24hrs clearly is too long.- Badges
- Science topic