Dear all,
I have antibiotics free stemcell culture. In the last passage, 2 days after I got bacteria contamination in all wells. I probably contaminated them during passaging because they were clear before. Since there were no antibiotics if they were contaminated I also should have seen them before.
Contamination resulted in cloudy bacterial propagation 48 h after passaging.
The bacteria look excessively strange theoretically they should be bacillus subtilis like bacteria, they are abnormally big like small worms ( of course I cannot carry worm contamination to cell culture). But worms do not create a cloudy view as well as they do not proliferate that rapid.
After I realize this cloudy structure, I washed each well 2 times, and I replaced the medium with the medium containing pen/strep+ Kanamycin (50ug/ml). My stem cells were already damaged so I wanted to see if I can eliminate them with this combination. Kanamycin is a very strong antibiotic at this concentration.
the images 48 h after passaging and 12h after washing is attached.
Probably Bacillus like bacteria gets in excessive amount it is due to the fact that it is resistance to both pen-strep and Kanamycin.
Now I will open a new stock that I think they are clean after I throw everything. I plan to start treating them with Gentamicin immediatly after they stick to plate. Do you have any other idea? And do you know what is this contamination?
Best Regards
Yağız
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