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How to tell my cells are contaminated? - (Jan/23/2007 )

I have stable cell lines of HEK293 cells with GFP fused with my protein.Since last week,lots of cells are floating in the media.Many cells are attached to the plate & growing fine.Today i saw something slimy in the media but the media was not cloudy.
Are the cells contaminated.
How can we detect contamination.
Do the cells grow fine if there is contamination.
Plz help.

-minnee-

QUOTE (minnee @ Jan 23 2007, 01:47 PM)
I have stable cell lines of HEK293 cells with GFP fused with my protein.Since last week,lots of cells are floating in the media.Many cells are attached to the plate & growing fine.Today i saw something slimy in the media but the media was not cloudy.
Are the cells contaminated.
How can we detect contamination.
Do the cells grow fine if there is contamination.
Plz help.


yes, your cells are contaminated, and they will not grow normally because the bacterias or fungus will grow quicker than your cells.
In addition, it is hard to get rid of contaminant.
Also, your bottle of media is contaminated.

You would better to thaw a new vial of HEK293 and make a new bottle of media.

Hope this help.

-Minnie Mouse-

cells do not grow fine with contamination. they usually end up very very sick, and then fall over and die.
try to get a new vial of cells. toss the media. clean the incubator. clean the water bath. clean anywhere you think might have caused the contamination.

V

-vetticus3-

If you have something very slimy could be a yeast (check by microscope if something round smaller than your cells and bigger than bacteria) then you are in trouble because is hard to get rid of it. I you found something slimy but can see the cause then is a bacteria. If you see something like transparent hairs then is a fungie. either case just stop all cultivation and clean with chloride or lysol (10%)all surface and rinse well. toss anything that could be responsible for the contamination, add an antibiotic to the medium. always clean the bottle of media with 70%alcohol and the must important, when working with cells never let a person stand next to you (2 reasons first is a distraction and you could miss an aseptic step and 2nd could contaminate the area if speak or touch something)

-merlav-