When growing human hepatocytes in culture, I have always added 1% penicillin/streptomycin to my DMEM/F12 media. I recently read that this may make my cells vulnerable to mycoplasma infection. Is this true? If so, why? Also, why do the antibiotics not affect the liver cells?
Ideally, you may grow cells without antibiotics, just it is much more complicated if you want to avoid the bacterial infection. It is easier to grow cells with antibiotics, that's why many people add it into the medium.
Ideally, you may grow cells without antibiotics, just it is much more complicated if you want to avoid the bacterial infection. It is easier to grow cells with antibiotics, that's why many people add it into the medium.
I have found that pen/strep is hard on human cells in culture (surprisingly), and opt for gentamicin instead! Its more stable, better spectrum, and less toxic for the human cultures we do.
We culture our hepatocytes always without pen/strep or any other antibiotics. By using good sterile working practice, this should not be a problem!
When you perform experiments with PenStrep in the medium, you can get some secondary effects because antibiotics can influence mammalian cells. As Sarah recommended, you should work in a clean environment to avoid bacterial contamination. Remember that most antibiotics do not kill bacteria, but only slow down their growth.
If you have to use antibiotics, run at least some experiments without as control first to establish that your cells are the same with or without antibiotics. Look at morphology, growth rate, viability etc.
Good luck
Sometimes, antibiotics are not only used to prevent bacterial contamination of the cell culture, but also to keep transfected features in the cells. But there may be some undesirable secondary effects (altered metabolism/susceptibility of the cell; baseline-stress) that may interfere with what you are measuring.
If I have the choice, I therefore prefer to use antibiotics once in a while (e.g. once a week/once in two weeks) for general culture maintenance, but do experiments with cells that have been passaged at least twice in media without antibiotics.
Good luck :-)
https://www.sciencedirect.com/science/article/pii/S0076687979581282/pdf?md5=ff87c86
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