2 replies to this topic

[meee]

Hello friends
I am working on HepG2 and my cells are contaminated, i use gentamycin (50microgram/ml) in media. the bacteria doent grow fast so to make media turbid but grows very slowly. I tried the same cells to grow media with penicillin, streptomycin, amphotorocin combination and without antibiotic. I am very scared of all these things.
So dear friends please do help.

Thanks

[klinmed]

meee, on May 3 2010, 07:38 PM, said:

Hello friends
I am working on HepG2 and my cells are contaminated, i use gentamycin (50microgram/ml) in media. the bacteria doent grow fast so to make media turbid but grows very slowly. I tried the same cells to grow media with penicillin, streptomycin, amphotorocin combination and without antibiotic. I am very scared of all these things.
So dear friends please do help.

Thanks

If you had a VERY precious (non-replaceable) culture I would suggest 100 ug/ml meropenem. It is antibacterial "kill all" I often use clinically. However, you should be able to maintain a simple to grow line like HepG2 without any antibiotic/antimycotic. Indeed, you should ALWAYS culture continuous cell lines without antibiotics. Antibiotic use just covers up bad technique. If your aseptic technique is so poor that you need antibiotic, learn correct methods from a competent colleague. Overuse of antibiotic can lead to mycoplasma problems etc. Also inappropriate antibiotic use can result in significant metabolic perturbations that can influence your experimental results. In short, DO NOT use antibiotics for routine culture of cell lines, you are just asking for problems

Hope this helps.

[leelee]

I agree with klinmed.

Throw your contaminated cells out and start again with a fresh batch.