Looking for an answer to this problem that hasn been affecting my cells for a while. Attached are example images of what Iv'e been seeing in my BV2 cell cultures for about a month now. Around half of all my coverslips have cells that have died after adding treatment (which can be either serum-free RPMI, or a treatment in serum-free RPMI), while the other half that recieevd the same treatment are perfectly fine. Seeding the cells and supplementing the media has not resulted in this effect, but only at the treatment stage does this ever occcur. Most of the time the entire coverslip dies, but occasionally I see this occur, where there is a clear border between living and dead cells. The timing of this effect is instant, as I have timed how long this takes to occur, and between adding the treatment and checking under the microscope (30 seconds apart) theyt have all died. I have changed out the media, treatment and plasticware several times and have relied on three different media bottles (RPMI-1640) with different LOT numbers. This has not changed anything. I think this may be ethanol getting into my media, but I have been extremely careful since the first instance and I don't see how this could possibly be happening.
Any assistance in answering this would be greatly appreciated.
The following RG discussion mentions BV2 and may help:
https://www.researchgate.net/post/Can_any_one_suggest_the_exact_protocol_for_NO_assay_using_Griess_reagent_Do_I_have_to_use_colorless_media_for_plating_the_cells
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