HI,
I am trying to recover frozen LUHMES cells bought from commercial source and encountered bacterial contamination twice on the 2nd day after thawing. I am confident that the contamination is not due to reagents used or my aseptic technique. I checked each reagent separately for signs of contamination, and I don't have any problem with other cells types I'm currently working with. Though I do it inside BSC, I suspect the contamination could be introduced during the coating procedure, where the protocol calls for drying the flask uncapped for few hours. Researcher already working with LUHMES cells, could you pls advise me on this issue. Thanks in advance.
Drying the flask should be performed in the BSC to prevent contamination. You can potentially dry the flask under UV light too. If you aren't already adding pen/strep to your media, you can try that.
Since LUHMES were immortalized using a v-myc construct with neomycin resistance, they are neo resistant and you can add neomycin or G418 (200 ug/mL) to the media to potentially kill any other contamination.
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