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Possible contamination - suggestions?


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6 replies to this topic

#1 csegal

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Posted 09 October 2009 - 07:07 AM

Hi,

I'm culturing adherent LNCaP cells and am concerned that I have contamination - see attached pictures. I contacted HPA, who I bought the cells from, and they suggested that it could be a fault in the plastic (no) or condensation on inside of flask (also no). Does anyone have any better ideas?

Generally, the objects are round, clear looking and larger than the cells. They are floating in the media at the level of the cells (i.e. not air bubbles at the surface).

I sterile filtered all the components that I added to the base media and started fresh reagents. The cells seem to be growing fine and the media doesn't seem to be exhausting quicker than normal.

Suggestions please?

Attached File  arrows_0121_cropped.bmp   417.65KB   122 downloads
Attached File  arrow_0129_cropped.bmp   511.02KB   74 downloads
Attached File  arrows_0127_cropped.bmp   250.06KB   74 downloads
Attached File  arrows_0123_cropped.bmp   261.97KB   69 downloads

#2 Dr Teeth

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Posted 09 October 2009 - 09:41 AM

Hi,

I'm culturing adherent LNCaP cells and am concerned that I have contamination - see attached pictures. I contacted HPA, who I bought the cells from, and they suggested that it could be a fault in the plastic (no) or condensation on inside of flask (also no). Does anyone have any better ideas?

Generally, the objects are round, clear looking and larger than the cells. They are floating in the media at the level of the cells (i.e. not air bubbles at the surface).

I sterile filtered all the components that I added to the base media and started fresh reagents. The cells seem to be growing fine and the media doesn't seem to be exhausting quicker than normal.

Suggestions please?

Attached File  arrows_0121_cropped.bmp   417.65KB   122 downloads
Attached File  arrow_0129_cropped.bmp   511.02KB   74 downloads
Attached File  arrows_0127_cropped.bmp   250.06KB   74 downloads
Attached File  arrows_0123_cropped.bmp   261.97KB   69 downloads



Are you sure these aren't dead cells?

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 mtrnbh

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Posted 09 October 2009 - 09:54 PM

Is there a risk of cross contamination with a non-adherent cell line?

#4 bob1

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Posted 11 October 2009 - 03:50 PM

dead cells.

#5 rhombus

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Posted 12 October 2009 - 01:47 AM

dead cells.



Agree with bob1, dead cells called "ghost cells"...where most of the intracellular material has been lost and a "ghost membrane" remains.

Rhombus

#6 eberthella

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Posted 12 October 2009 - 03:06 AM

Would a live/dead stain help?

#7 csegal

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Posted 12 October 2009 - 03:15 AM

Thanks for the suggestions.

I'm pretty convinced it is not cross contamination with another cell line as I've just purchased these from HPA and I'm not culturing any other lines (and I have sole use of the incubator). As for ghost cells, I thought I knew what these looked like and these objects don't look like what I had thought were ghost cells! I'll do a stain and see what happens.

Thanks for the input everyone.




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