6 replies to this topic

[TimGG]

After a long weekend, I found out my CHO cell line are all dead inside the humidified chamber, try to figure out why they all die!!

The media was cloudy pink for all 3 T75 flask.

2 of those I was spilt on last Friday, and 1 i spilt on last Thursday, so I figure if there is something wrong during the spilt, it should only happened to one set of cell, but it didn't~~ they all dead......

The CO2 gas i have been using for a month and nothing happened before.....

So if anyone have any idea why this happened, i'm pleased to hear~~~

Thanks a lot.

[gfischer]

Put some medium in a flask/plate and put it in the incubator for a couple days. I suspect that your media is contaminated.

[TimGG]

gfischer, on Oct 28 2009, 01:17 AM, said:

Put some medium in a flask/plate and put it in the incubator for a couple days. I suspect that your media is contaminated.

I've tried it for three days and nothing growth on my media, any more ideas from anyone?? I just trying to avoid this happen again~~~

[gfischer]

TimGG, on Oct 31 2009, 09:42 AM, said:

I've tried it for three days and nothing growth on my media, any more ideas from anyone?? I just trying to avoid this happen again~~~

Random contamination happens occasionally, usually due to a one-time lapse in aseptic technique. Just to be safe, I'd switch to new batches/aliquots of as many components (media, serum, growth factors) as you can without bankrupting the lab, and continue as normal. You may want to be a little more conscious of your technique.

[TimGG]

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It happened again~~~

I start to wonder is this a mycoplasma infection, I have attached some photos and hope if someone have experience in this can share opinion on this.

i start to search for mycoplasma detection kit and drug to cue this infection, and it would be great if we can get it from Sigma-aldrich.

Thanks for the help.

[bob1]

It's definitely bacterial and very highly contaminated, you should be able to see the bacteria under high magnification - if it was yeast you would be able to see the cells at x100 mag, other fungus usually appears as filaments.
It is unlikely to be mycoplasma - you are very unlikely to get visible levels of mycoplasma in a culture. You are better off throwing out your cultures and starting again than you are treating the cells for the contamination. This is the case for any contamination, as the treatment agents usually suppress low level contaminations that will still affect your results.

[madrius1]

Exactly. Throw everything away and start with fresh media.