Mycoplasma - input needed! - Please help me by answering these questions! (Jul/10/2005 )

-- What are your common lab practices for avoiding mycoplasma infections?
-- What do you do when you have it? For both your lab at large as well as the cells themselves?
-- How do you commonly maintain your incubators?
-- Have you ever experienced mycoplasma contaminating other vials in a common liquid nitrogen tank?

Thanks in advance!

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[quote=MaximinaNYC,Jul 10 2005, 10:14 AM]


Hey Max,

to answer your questions:

-- What are your common lab practices for avoiding mycoplasma infections?
we have separate TC flow hoods for mycoplasma free and "possibly contaminated" cells. Also we test all cells when being set up for the first time and every 3 months.

-- How do you commonly maintain your incubators?
Actually we do not do much. Surprisingly, and this may sound weird, but everytime we clean out our incubators, we get some sort of yeast or other extra terrestrial bugs growing together with our cells. We have a semi-annual tc room clean up.

-- Have you ever experienced mycoplasma contaminating other vials in a common liquid nitrogen tank?
You know, i hear this a lot and frankly I dont believe it. Its highly improbable, although i concede not entirely impossible, that some mycobug swims merrily through the N2 pond to its neighbour. Hmmmm

Anyway i hope this helped. I hope others will also share their experiences..


take care

Sjon

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we do nothing in particular except testing regulary our cells (about every 3month and testing new incoming cells


We never experienced mycoplasma. But in a close lab, they just put eveything in the washer and after spray all the cell culture room and stuff with a commercially available spray. That worked

once per year, we bring them at 95° 1h and that's all


I don't believe it's possible. Freezed mycoplasma is not movable... But i assume that if you freeze a vial with mycoplasma inside or sticking it, the same "mycoplasma culture" can re surface month laters... To get rid of that, you can spray all tubes you get out from the nitrogen briefly after thawing.

fred

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Hi,

We are probably a bit lax compared to the others, we test our cells on thawing and then every 6 months.

When we have experienced mycoplasm in the past we threw out the cells and started again, unless they were precious cell lines which we treated they do clear up with a commercial removal agent but I always worry if they will ever be 'normal' again. The incubators, hoods etc. were disinfected by 'bombing' overnight. Set up absolute myco free incubators and hoods and adhere to them strictly.

Every six months we 'bomb' the incubators (not exactly sure what is meant by that our tech does it).

As for contaminating in liquid N2, I would say absolutely impossible. Not only would the myco have to survive liquid N2 (which it does) but would have to enter sealed vessels.

Scott

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work as clean as possible


test every now and then, roughly every 3 month, or when cells start to behave stragely in standard tests....
we throw the cells away, thaw a new batch, test that and go on...

we have a copper plate at the bottom of our incubators, each incubator gets cleaned thoroghly one every six month (treated with disinfectants/fungicides etc..) and once every two years they are disinfected by formalin treatment (or so I'm told)



Never had it, but I do think it's possible, though unlikely

mike

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to get rid of mycoplasma we tried ciprofloxacin as supplied by sigma aldrich (its cheap.....20 Euro for 5 mg)....we used a mycoplasma-contaminated huh7-line and the cells were cultivated with and without cipro for 2 weeks.....after that ,our test (enzymatic...dont no how it works) said, that the treated cells are free of this little bastards.....

now i will cultivate them for 2 weeks without cipro to look, if the mycos are gone forever or will come back.....


Greetz

K_R

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we treated the cells with cipro at first, too, but found that cipro itself changes some (immunological) properties of the cells like senitivity to stimulants and the like, so we stopped it and threw the cells away...

mike

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