Contamination issue in cell culture - (Nov/16/2004 )
Hi... like every suggestion's in this forum.. if any contamination happens, just draw out your flask, and start with the new one...
your problem is the same with mine, few months ago.. but it was too tiring wait until your cells proliferating, i think its kind of waste of time... and because i need to test the performance of biomaterials, the cell proliferation data seems not reliable if you use "sick" cells.. so, if you have any other vials, just start it with new cells... and dont forgot to clean your incubator too before start with these new cells... hope it works
Hi... like every suggestion's in this forum.. if any contamination happens, just draw out your flask, and start with the new one...
your problem is the same with mine, few months ago.. but it was too tiring wait until your cells proliferating, i think its kind of waste of time... and because i need to test the performance of biomaterials, the cell proliferation data seems not reliable if you use "sick" cells.. so, if you have any other vials, just start it with new cells... and dont forgot to clean your incubator too before start with these new cells... hope it works
Thanks for the tip
Good suggestions all, and I would concur - if you have a yeast infection, or any infection for that matter, you will be best served by destroying the infected cells and sterilising the incubator.
I know this is very frustrating, especially if the infection occurs in a recent passage which you are experimenting with, but it is a sad fact of life in micro labs.
Hi... I have a question. Unfortunately our Bioreactor got contaminated with some sort of bacteria. It made an awful odor in the incubator. Is there anyway to get rid of that smell?!
.l Mine are MA-104 clone cells infected with the parasite Neospora, and since we obtain the isolate from mice to further adapt them to in vitro culture I guess some sort of contamination is coming out of this animal model. At the beginning we didn't figure we had this contamination because we usually work with medium containing Pen-Strept-Fungizone, but now we try to keep cultures without antibiotics and we have started to see at this tiny "bugs" with irregular shape, either alone or glued each other and sometimes adhered to the cells. 
At first, cultures look fine and medium has the appropriate pH but suddenly they raise their number and pH drops to 5.5 (amazing yellowish medium), so I must get rid of flasks before contamination affects the rest of what we keep on the incubator. We have run several tests (agar and brot culture, PCR for mycoplasma detection) and nothing shows up. If someone looks at the video and photo we are attaching, I would appreciate any advice you can give. Thanks!!! 
HALLO,
you are not alone.
please look at photo from HSC primary culture... 1 week...
greetings
hi...
this seams like a really bad contaminat, and I got it too, does somebody deal with it?
everyone is just complaining but does somone win? I will like to know how to deal with this parasite?
Desperate begginer
I have experienced the same as most of you: Black wiggly rods/spheres that slowly propagate in media without turbidity or media color change. They aren't observed until high mag, ~400x, and can then be seen as very small compared to cell body (of A549, 293T, CaCO2, Hela etc). They are Pen/Strep resistant and slowly multiply. In my A549s they seem intracellular and on surface of cells, but also extracellular (as is the case with most of my other cells). I don't believe this to be 'brownian motion' or other lot. While people's suggestion of "Good Technique" is noteworthy, it's about as useful as telling someone don't fall down after they're already on the ground. All my stocks are contaminated, there is no going back. I have 4 years of research at stake. I NEED to know how the hell to get rid of them, not a slap on the back and tell me to be more aseptic. I will post some pictures because the ones I'm seeing posted look kind of poor quality so I will try to point them out etc.
I have been doing cell culure for around 30 years now and have NEVER been able to clean up a contaminated primary. When I was first in the lab I tried washing 10/20 times the monolayer with PBS, adding extra Pen/Strep etc. IT NEVER WORKED, the contamination always came back. My collegues at the time said throw the cells away and start again. Use better TECHNIQUE to start with to reduce the chance of contamination. I of course did not listen to them and tried anyway. Try multiple washes and adding extra antibiotics if you want. When the contamination comes back you will say to yourself ... " Why did I not invest that time in isolating fresh cells.
The other thing to remember is WHAT DOES THIS DO TO THE CELLS. Will their morphology, phenotype, biochemistry, receptor expression CHANGE BY DOING THIS.
Just wanted to feed this very valuable topic.
I never had a contamination in the short time i've been working on cell line.
But.... we work with three different ppl on the same flow so we are monitoring and helping eachother.
Here is what I found out:
* Using 70% Ethanol (Desinfectol) matters. A colleague of me just though naaah, bullshit i'm using 100% Desinfectol (wich is actually 95%Etoh).
He got contamination. Why: above 70% the alcohol dries too fast. In time you rubbed the whole surface of your hands/gloves the alcohol is damped out already.
The same goes for cleaning your flow.
BTW: This colleague has been doing cell culture over a year.
* If you suck off the liquid medium when refreshing make sure to cleansen the tubing. You can do this by sucking off 5% RBS liquid as last step.
* Also don't use antibiotics. -> avoiding Resistant strains.
Hope those hints help.
"* If you suck off the liquid medium when refreshing make sure to cleansen the tubing. You can do this by sucking off 5% RBS liquid as last step."
or you could just squirt the ethanol up the tube