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Mycoplasma Testing in culture medium - (Nov/05/2005 )

Hey,

I've just started my PhD and I'm new at cell culture.I set up cultures of MCF-7 and T47D cells from liquid nitrogen, they grew just fine, I split them and about 4-5 days later... the culture went cloudy and murky and i could see little (very tiny) bugs wriggling around in the culture. I thought that a bacterial infection normally appeared much quicker than this? So I have put up flasks containing just culture medium, and so far (5 days later) the medium still looks fine. Is it likely that the cells were infected when they were frozen down? I don't think it's a mycoplsma infection as the cells grew very quickly to start with, but is it possible to do a hoechst stain on just medium?

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-HannahBelle-

that could certainly be a bacterial infection

could have happened when you split them, or changed the media afterward?

sometimes they grow really fast, sometimes not...depends on the type of bacteria and what is in your media.

it is my understanding that mycoplasma is not really 'visible'; there is no cloudiness or turbidity, it is hard to detect because mostly you just see morphological changes in your cells.

good luck

-aimikins-

hi
just pipett 200┬Ál of supernatant and do a PCR test for checking mycoplasma...

do you use peni/strepto in your medium? if yes you have sthg resistant to them. So your cells aree lost...

-fred_33-