Last week I had consecutive contaminations of some of my mammalian primary cells alone. It started with the cultures turning turbid after a day or two of incubation. I wiped down the entire laminar hood and sterilized the incubator along with changing its filter. I kept defreezing new vials of cells and then again there were rod shaped structures that didn't allow cells to grow. I observed the medium alone under microscope but it appeared fine. Today I found these spore shaped structures (under 20X objective), not rods. But these didn't seem to disturb the actual cell culture. Could these be bacterial contamination? If so could they change shapes?
It looks like yeast to me, which can bud to look rod like. Rinse multiple times with sterile buffer and add Amphotericin B to your culture medium to ~1 ug/mL. Treat for 1-2 weeks.
You could have the contamination in your frozen vials. In this case either obtain a new stock (preffered) or treat with amphotericin B as above. In case it is yeast this will help.
the structers were comproved bacterial spores ? Try the Gram stain.
Thanks all. I suspected the frozen vials to be contaminated as well and kept defreezing new ones (but of the same mother stock). The contamination kept reappearing until I threw away the medium and used a newly prepared one. I haven't seen anymore familiar looking particles in my cultures till now, yet my cells do not grow well. I will give it sometime to see how it goes.
Is it ok to treat cell cultures with anti-fungal? Will it affect my experiments in any way? Just asking as I have never done this before nor seen anyone I know do it, except for adding the anti-bacterial strep/pen.
Thanks again!
Hi Anusha, you should not use your cells while they are contaminated or unter treatment and I would give them a recovery time afterwards. For me this would be the last resort if you absolutely can't get a new batch of cells.
We have not done it for fungi yet, just for mycoplasma. Are your cells checked for mycoplasma? this could be a reason why they don't grow properly even though you don't see a contamination.
Hi Victoria,
I am suspecting mycoplasma as well, as am running out of ideas. But do they also act in a way as to not let cells attach to the surface and grow?
Thanks!
That could be one possible consequence. The effects from mycoplasma can be quite diverse and inconclusive (from detaching to killing and even more proliferation).
Only a test for mycopolasma will give you clear results
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