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Stringy rapidly growing contaminants in primary culture

mice fibroblasts

Best Answer SusieQ, 07 July 2014 - 10:14 PM

Hi Ahyi,

 

I've seen these deposits before, but I've never had the experience that my cells started dying or anything out of the oridinary, so after intricate thinking and discussing it with my coworkers, I dismissed them as dust or debris (perhaps unjustified). I myself have no experience with primary cell culture, however, here's my two cents:

 

- Is your medium discolored and/or cloudy? This may incidate a contimatination of some kind (bacterial, most likely).

- How much serum do you use in your medium, if at all? I've been told and I've also seen for myself that, sometimes, high percentages of serum can cause coagulated protein deposits, looking like tiny strings or ribbons. I've no clue as to whether it's a certain kind of protein that causes this, or whether it's protein at all, but I got rid of this by filtering the (horse)serum before adding it to the medium. It didn't seem to harm my cultures afterwards.

- Does your colleague from whom you received that second batch of cells have this kind of problem as well, or do any of them know what this can be?

 

If you want to be sure whether it's a biological contaminant you can remove the medium that contains this phenomenon and reincubate it in a clean flask without cells. If it's then still propagating, it may be an organism (if you want to be really really sure, you could do a nucleus stain, like DAPI). If it's a protein deposit originating from added serum to the medium, these can already appear after you've warmed up your medium. You can see them if you hold the bottle up against the light, they're kind of floating around.

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15 replies to this topic

#16 Elaine123

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Posted 27 June 2015 - 02:27 AM

These stringy substances are all over the flask (not just a couple). Their length was as long as the image above but there was some that were very short as well. Also they were seen in different flasks with different cell trains. 







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