I just want to know whether it is necessary to add antibiotic to cell culture media?
We use 1% antibiotic and antimycotic solution to prevent bacterial and fungal contamination.
This solution contains 10,000 units/mL of penicillin, 10,000 µg/mL of streptomycin, and 25 µg/mL of Fungizone (amphotericin B)
The antibiotics penicillin and streptomycin prevent bacterial contamination of cell cultures due to their effective combined action against gram-positive and gram-negative bacteria.
amphotericin B prevents fungal contamination of cell cultures due to its inhibition of multi-cellular fungus and yeast.
In animal cell culture I add 100µg of penecillin and streptomycin. Its only necessary if you dont ensure you cant keep the culture clean.
It will make your culture free from contamination, so better to use it though its not necessary.
Nothing is necessary. I know many labs that don't use any at all. You can buy standard 100x Pen Strep from almost anywhere (including invitrogen). We also use gentimycin (200ul in 500 ml) from Invitrogen for cells coming directly from humans or for hard to kill bacteria.
1% pen-strep is commonly used as antibiotic system in cell culture.
If you ask whether is it necessary to add the antibiotic, it depends on what you are more comfortable with.
Some people prefer to add the antibiotic to prevent bacterial growth in their cell culture, so that they can have peace of mind.
Other prefers not to add any antibiotic so that they can immediately detect any bacterial contamination (especially in the CO2 incubator) and swiftly deal with the outbreak.
Keep in mind that by adding antibiotic, you are just suppressing the growth, it doesn't necessarily mean that the bacteria are not there...
Hope that helps.
I never use it as makes you sloppy in asceptic technique. Also depending what you are working on it can also interfere with your results. Best to try your cells without antibiotic and if grow fine and show no signs of contamination then dont use it.
One more suggestion: if you are using primary cell cultures then I will suggest to use it, as there is high probability of contamination.
The recommended concentration is 100 IU/ml of Penicillin and 100ug/ml of Streptomycin.
Rathish Rajarathinam thank you sir for your kind repply
But How we calculate100 IU/ml of Penicillin for 100 ml of Media
Matthias Scheb-Wetzel., Abhijit Shinde, Peter Gould, Kai Lun Chang, Kathleen Ignatoski....Thank you all for your valuable suggestions and opinion...
The concentration of penicillin is per ml, so calculate accordingly. Also, the antibiotic solutions are available as 100X stock solutions from Gibco and from other manufacturers. You could buy one and use it which will ease your work.
Hope this information helps you!
I agree with Peter, it is better to have good aseptic technique and do without antibiotics but there are times when they are necessary, particularly when initiating primary cultures. We routinely use 1% pen/strep (Gibco, 15140) in our primary cultures throughout the life of the culture, however some people prefer to switch to antibiotic free medium after the first 7DIV.
Other factors to consider are the cleanliness of your culture lab; the number of users and if you suspect other peers or students may use your reagents!! The more users the more likely bacteria will be brought into the lab. If you have a well-equipped and maintained lab and your aseptic technique is good then technically antibiotics should not necessary when culturing cell lines. Perhaps trial a few cultures without antibiotics and see how it goes. Good luck.
We add antibiotic-antimycotic from Invitrogen to all our cell cultures. we do large volumes eg 50L... hence we cant risk contamination. As Karen mentioned it is better to avoid them by having good aseptic technique and keeping the hood clean and trafic to a minimum
We use 1% antibiotic and antimycotic solution to prevent bacterial and fungal contamination.
This solution contains 10,000 units/mL of penicillin, 10,000 µg/mL of streptomycin, and 25 µg/mL of Fungizone (amphotericin B)
The antibiotics penicillin and streptomycin prevent bacterial contamination of cell cultures due to their effective combined action against gram-positive and gram-negative bacteria.
amphotericin B prevents fungal contamination of cell cultures due to its inhibition of multi-cellular fungus and yeast.
Kunal Pal, you copy pasted Nimal Raveendran's answer yet again. Is this your best skill?
What do you think might happen if 100x of the antibiotic solution is added to the flask where contamination is observed?
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