Hi,
I am culturing adherent SF9 cells in SF900 II medium (supplemented with 10 % FBS and penicillin and streptomycin antibiotics) at 27 C.
My cells grow perfectly till 2 passages. However, in the third passage, it gets contaminated with the fungus. I have tried using everything fresh (media, fresh revived cells, wiping the incubator and laminar hood, using different laminar and culture rooms, etc.). Can anyone explain what the problem can be here? and provide some tips to prevent the contamination? Thanks in advance.
Fungal contimination is difficult to get rid of.
If you are still having issues when switching to a hood and room where other's didn't have a problem... It is almost likely somehow you are introducing it during the passages... If you can, see if anyone else can passage your cells without any issues and work backwards from there.
You can try adding an anti-fungal (like amphotericin B)... and see if that helps.
Are you sure it is fungal infection? SF9 medium is prone to form precipitates that look like contamination, if the surrounding atmosphere is pretty dry.
DearJulijus,
First I recommend you, if there is someone else working on the same cell type or same medium, ask her/him to culture instead of you.
Or you can borrow some material for culturing.
second, I recommend you to change all of your pipets.
and finally check your 70% alcohol.
you know when you unfreeze that cells, be careful so much.
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