Recently, in my primary culture of placental epithelial cells unwanted structures of microbes have appeard. Medium was cloudy and changed the color to yellow during culturing. I tried to treat my cells with amphotericin B (0,25 microgram/ml of full-suplemented medium) - I changed fresh medium every 2-3 days, but it didn't help and after 3 weeks infection eliminated eligible cells. Could you give me an advice what kind of microorganism (I suspect some fungus) is it? And whether it can be removed from culture of animal cells in the future? How can this be achieved?
Hello Monica
In my opinion it seems that you have contamination with either yeast or candida. I do not have experience with primary placental epithelial cells, but we do use primary hepatocytes which we infect with mosquito material (basically super dirty with an assortment of fungi!). In order to contain fungal infection we use Fungizone (also Amphotericin B) at a working concentration in the medium at 1.25 ug/mL. This works quite well for us, as it does not disturb the primary cells. If you can run a quick IC50 of Amphotericin B on your cells, and then if possible use a higher concentration, you may control the fungal contamination.
Good Luck
Best
It seems to be yeast contamination. I would advice you to use penn/strp/amphiteracin. Throw away these cultures and clean the incubator and other areas properly.
Thank you all for helpful observations and advice.
I normally use Pen/Strp solution at a concentration 100 IU/ml and 100 µg/ml, respectively. After I had noticed contamination (cloudy culture medium in the flask), I started to use amphotericin B (0,25 ug/mL) additionally. Cell culture was treated for 3 weeks, but it was getting worse. Finally (yesterday) I decided to throw away this flask (because only one, among 4 after isolation, was contaminated).
I'm worried about the other cells, although so far I've not seen suspicious yeast cells. I've washed UV chamber with Lysoformin. What more should I do in this case? How to clean the incubator? What more should I do in this case?I have a small amount of antibiotic/antimycotic solution (with 10000 U Pen, 10 mg Strp and 25 μg amph B/ mL). Should I add this solution to remaining "healthy" cell cultures and at what concentration?
I will be very grateful for any suggestions.
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