I extract the primary RPE cells from pig. As I see in several time extraction high contamination of bacterial in my cell culture flask in the 3rd day after extraction. Sometimes they survive untill one week but finally high bacterial contamination detach and kill cells. I wanna to increase the antibiotic dose (more than 2%). I use usually 2% pen/strep in medium. Does anybody know to How much I can increase it without showing toxic effect to the cells?
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The tissue must be placed in a solution with 200 U/ml of penicillin, 200 μg/ml streptomycin and amphotericin B 50 μg / mL. After isolation of the cells, they are incubated in a medium containing 100 U/ml penicillin, 100 μg/ml streptomycin and amphotericin B 25 μg / mL.
In my work, we use serial dilutions from 5 μg / mL to 500 μg / mL to avoid cytotoxcity by increase concentration by the tested drug.
Hi Elmira,
It's been a while since you brought up this question, hope everything goes alright now. I think it's fine that you go up to 5% of the antibiotics you mentioned in a relatively short period of time, with anti-fungi drugs added. Besides, it might be helpful to rule out the mycoplasma contamination as well. Also, changing the media more frequently may help.
Best,
Yi
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