Opinion: Is my culture contaminated (yeasts)? - Pictures includede (Mar/13/2010 )
Hello everybody,
My brand new neuroblastoma cultures seem doomed to tragedy. I'm not familiarized with these cells but I found refringent ovals all dispersed in the culture and attached to the surface of the cells.
Medium is RPMI with 10% serum, 1% L-gln, 1% penicillin-streptomicin and 2,5 ug/ml of fungizone!!!!
Medium is replaced dayly.
I can't throw these cells away!!
This is a panoramic view of my cells:
No chains of budding can be found but look at this weird formation that I found in one of my plates:
Besides that, I've observed that when the cells grow at low confluency, they tend to aggregate and form a kind of tumoration, but I don't know if this forms part of the contamination process.
What do you think?
It is a bad idea to culture with antibiotics and antimycotics (antimycotics are worse - they affect your cells a lot as well as the fungi).
Your cultures look pretty confluent to me - lots of cell process touching, which is usually enough for neural cells. It is pretty hard to tell whether you have a contamination or not - using antibiotics and antimycotics pretty much ensures that you don't, though there could be a cryptic infection inside or attached to the cell.
To test - remove the pen/strep from a culture, leave for a couple of days, repeat with the fungizone. If you get obvious contamination appearing, then there is a problem - throw the cultures out and get some fresh stocks in and culture them properly!
EGF on Mar 13 2010, 04:34 PM said:
My brand new neuroblastoma cultures seem doomed to tragedy. I'm not familiarized with these cells but I found refringent ovals all dispersed in the culture and attached to the surface of the cells.
Medium is RPMI with 10% serum, 1% L-gln, 1% penicillin-streptomicin and 2,5 ug/ml of fungizone!!!!
Medium is replaced dayly.
I can't throw these cells away!!
This is a panoramic view of my cells:
No chains of budding can be found but look at this weird formation that I found in one of my plates:
Besides that, I've observed that when the cells grow at low confluency, they tend to aggregate and form a kind of tumoration, but I don't know if this forms part of the contamination process.
What do you think?
It would be very useful to see "phase contrast" images of your cells/contaminant. In my experience it is virtually impossible to identify possible contaminants using bright filed images.
As bob1 rightly states "it is a bad idea to use antibiotics and antimycotics" in regular culture. These compounds are extremely dirty and WILL effect cells in lots of different ways.
Again in my experience it is impossible to completely eliminate yeast infections. Yeast infections are easy to see as they proliferate quickly and have a very distinctive shape i.e. a sphere with a bulb at one end
Phase contrast images will help us alot
I hope this is useful
Kindest regards
Rhombus