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Cell culture contamination


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#1 CFLM00

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Posted 15 October 2009 - 02:48 PM

Hi
Two weeks ago I received a cell line from ATCC. The proliferation of the cells was very low and early I suspected that something was going wrong. I saw With time (after one passage) the cause was revealed and the intruders showed themselves (attached photos). The medium did not turn yellow and almost no turbidity was seen. Can anyone help me to identify the outsider? Should I try a contamination removal? If so, how?
Thank you in advance

Attached thumbnail(s)

  • Attached Image: SNAP_105906_0004.jpg
  • Attached Image: SNAP_110224_0006.jpg
  • Attached Image: SNAP_105647_0002.jpg


#2 LostintheLab

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Posted 15 October 2009 - 05:03 PM

If you got this cell line from the ATCC, then start with them first; get in touch and tell them the problem.
You will have to fill out a form about your media, culture conditions etc. etc. (so make sure your media is NOT the problem! Put the media in an empty flask and culture without cells to check and see whether its contaminated).
The ATCC will probably send you another batch of cells free of charge assuming the fault lies at their end and they'll certainly help you out if you have problems.

btw- It looks like a fungal infection most likely, I wouldn't worry too much about what contamination it is, more that it is contaminated.
Check your incubator and hoods are clean and that your media is clean and get in touch with the ATCC.

Edited by LostintheLab, 15 October 2009 - 05:05 PM.

I knew it! I knew it! Well, not in the sense of having the slightest idea, but I knew there was something I didn't know.

#3 CFLM00

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Posted 16 October 2009 - 02:34 AM

View PostLostintheLab, on Oct 16 2009, 02:03 AM, said:

If you got this cell line from the ATCC, then start with them first; get in touch and tell them the problem.
You will have to fill out a form about your media, culture conditions etc. etc. (so make sure your media is NOT the problem! Put the media in an empty flask and culture without cells to check and see whether its contaminated).
The ATCC will probably send you another batch of cells free of charge assuming the fault lies at their end and they'll certainly help you out if you have problems.

btw- It looks like a fungal infection most likely, I wouldn't worry too much about what contamination it is, more that it is contaminated.
Check your incubator and hoods are clean and that your media is clean and get in touch with the ATCC.



Thank you for your answer!
I contacted ATCC, but as was expected they don't assume any responsability. They "do not routinely replace cells for reasons for contamination unless there is a reason to suspect that there is a problem with the cells which originated at ATCC". I need to order a new vial....
I did controls of everything, including medium and I didn't find any signal of contamination! None other cell line handeled in the same hood and incubator (and cultured in similar conditions) in our lab showed to have intruders....
Anyone have knowledge with similar situations with different outcome





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