I am working with MS1 endothelial cell line (mouse), and have noticed a recurrent contamination which appears after day two in culture. I tried again by changing to new media, new trypsin etc. But still noticed the culture got contaminated. So I am suspecting the original cryo-stock to be contaminated.
My question is that what is the most common type of contamination associated with these cells, and how to eliminate it? It's getting to the point where I am really frustrated, since I couldn't proceed in my experiments!
Thanks in advance!
I definitely agree with Thomas. It sounds like a contaminated cryogenic-stock and would be less complicated (and less expensive in the long run!) to start with a fresh cell line. It is always "hit and miss" when relying on the quality of frozen cell lines, especially if you were not the one to prepare them originally.
Thank you Thomas and Christine,
I also suspect that the stock is contaminated, and try to obtain fresh cells for my new experiments.
I would also review your cleaning procedures, just in case. In my lab we clean every 3 months or so with a surface disinfectant (the cabin, including its walls and behind the working surface; the bath and the incubator). For maintenance we first clean with 4% benzalkonium chloride, and then 70% ethanol.
In the bath we use autoclaved distilled water and Aquaclean (a microbiocidal additive for baths); in the incubator water we put copper sulfate. We also avoid talking when opening the incubator.
I hope it helps.
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