Hi everyone!
I am trying to culture cells from ATCC (ref: CRL-1520-ATCC) in order to obtain the A2B5 antibody. As the company recommends, I use a DMEM formulation from Sigma-Aldrich (D-7777) with a sodium bicarbonate concentration of 1.5g/L.
I did not use antibiotics as it was not specifically recommended and it was the first time of culture so no contamination was expected. However, after two weeks of culture the cells were full of bacteria.
Does anybody know the best way to culture this cell line? Has anybody used any different media from the ATCC DMEM?
Thank you in advance!
It should be fine just to add antibiotic/amtimytotic to what you are running, something like http://www.sigmaaldrich.com/catalog/product/sigma/a5955?lang=en®ion=NZ
If you don't want to use antibiotics you have to be very stringent with your aseptic technique regardless of what media you use.
Hi,
Obviously it is not a medium rather handling issue. Like Daniel has suggested, care should be excercised while handling the cells. In my lab use of antibiotics (other than those that are used to establish stable cell lines) is a crime and in past 5 or so years I do not recall any major incidence of culture contamination. In fact it is a good practice to get ones self trained in culturing cells without antibiotics for it pays off in longer run.
Hi
You must use antibiotic in recommended dose. i am agree with above said suggestions. Few more suggestions are antibiotics should not be used routinely in cell culture, because their continuous use encourages the development of antibiotic resistant strains and allows low-level contamination to persist, which can develop into full-scale contamination once the antibiotic is removed from media. Further, some antibiotics might cross react with the cells and interfere with the cellular processes under investigation.
Antibiotics should only be used as a last resort and only for short term applications, and they should be removed from the culture as soon as possible. If they are used in the long term, antibiotic-free cultures should be maintained in parallel as a control for cryptic infections.
Hope this helps little
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