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Thread: Cross contamination in cell culture...

  1. #1 Cross contamination in cell culture... 
    Forum Freshman RathDinen's Avatar
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    Good day everyone

    Do anyone know how to detect cross contamination in culture? especially contamination with Hela cells...

    Thanks


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  3. #2 Re: Cross contamination in cell culture... 
    Moderator Moderator TheBiologista's Avatar
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    Quote Originally Posted by RathDinen
    Good day everyone

    Do anyone know how to detect cross contamination in culture? especially contamination with Hela cells...

    Thanks
    I'm sure there's probably a number of standardised protocols for this.

    What cell type is your culture supposed to be and what equipment/techniques do you have access to? HeLa cells have more chromosomes than normal human cells, but some other cell cultures will display that too. You might be able to stain for some specific cell surface marker and do FACS.


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  4. #3  
    Forum Freshman RathDinen's Avatar
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    I'm using MDA MB 231, human breast cancer cells...
    I just doubt that the cell i culture is really MDA MB 231..
    The morphology under light microscope seems like Hela...
    I did comparison with the cell's image from the ATCC but it just getting me confused...
    Anyway, thanx for ur suggestions... :-D
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  5. #4  
    Forum Cosmic Wizard spuriousmonkey's Avatar
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    There was a study showing that most cell lines used in labs aren't actually the cell lines that they were supposed to be.

    So you shouldn't be too surprised.

    I think you are on the right track to check your cell lines. All your experiments in the future (and past) could be a waste of time if you didn't.

    I'm not a cell guy, so unfortunately I do not have a clue how.
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  6. #5  
    Moderator Moderator TheBiologista's Avatar
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    Ok, what I gather from some light reading is that you want to do PCR for HPV-18 (human papilloma virus) DNA. As far as I can tell, the HeLa line was immortalised by transformation using that virus so you should be able to find HPV genes in there. If you have access to a pure HeLa culture, use that as a positive control.

    Of course if your cell line was also transformed in that same way then you have a problem. Possibly you'll have to try figure it out by morphology. In that case I'd start by doing flow cytometry and just look at forward and side scatter to see if you have two morphologically different populations in your culture (assuming HeLa hasn't taken over entirely which it might have). Again use a pure HeLa culture as a control.
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  7. #6  
    Forum Masters Degree samcdkey's Avatar
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    The easiest way I can think of is to run a DNA fingerprint gel with pure cultures as standard markers [you can run just the MDA MB 231 or in addition, a lane with a pure Hela culture]. If there is more than one cell line you'll see it in the bands.

    The difficulty is in getting the "pure" culture for the standards. :P
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