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Another contimation problem

Another contimation problem - Cell Biology and Cell Culture

Another contimation problem - Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology.

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Old 03-17-2010, 01:19 AM
Pipette Filler
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Default Another contimation problem

Any help would be greatly appreciated. This has been a problem for about 1 or 2 years now and we are getting desperate! To start off, I'll give everyone some background. I didn't join the lab until 2 months ago (after all this happened) so I will try my best to explain.

We ordered a rare primate cell line from Coriell and went about culturing it. However, a strange hair-like particle started appearing in the media of all the cells (293T, CHO, etc). At first, we didn't suspect that it was a contamination problem but it was consistently found in all our cultures. Finally, after some investigating, our lab contacted Coriell about this problem. They claim that they didn't see anything in their cultures and agreed to send us another flask, free of charge. The older lab members have told me that as soon as they received the flask, they put it under the microscope before unsealing it and saw the same exact thing. Again, Coriell didn't acknowledge the problem.

To completely decontaminate the tissue culture room, EVERYTHING was thrown out and the hood was dismantled for disinfection. There was a theory that this thing was in the incubator so we even got an elaborate new incubator that cleans itself and threw the old one away. We put in hepa filters all throughout the lab. Everything was replaced and put back. The first day that the tissue culture room was up and running, we found it on the outside of the flask. Before long, it started appearing in the media again.

I started doing work in the tissue culture room after it was decontaminated. I found it in all my cells, especially in the ones that I am performing antibiotic selections on. I don't see more than 1-3 in 1 flask so it doesn't seem like it's getting wildly out of control but it is still stressful because I don't know what effect it has on my cells. I have done a lot of cell culture work before joining this lab so I know that it isn't my technique. I've started using a pen/strep/antimycotic solution in my media and I don't see it before I passage but as soon as my cells are split and new media is put on them, I see one in there. However, it disappears before it is time to passage again. I wipe everything down with Vikron and then 70% ethanol before putting anything back in the incubator. I have 6 cell lines going and I will have 5 more soon.

We really want to get rid of this problem and we want to find out what it is exactly. I have attached a few pictures of this thing. Please help us pinpoint what it is!! Our last resort is to hire a company to come out and decontaminate the whole room using a hydrogen peroxide vapor generator. That will be REALLY expensive so obviously we want to get rid of this with a cheaper solution.

Using our brand new awesome microscope and camera, I've taken some pictures. Any input would be greatly appreciated!!! Please help us figure out what this is. Sorry for the long post but I thought it would be best to include as much information as possible. Thanks!
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Old 05-17-2010, 09:53 AM
Pipette Filler
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Default Re: Another contimation problem

ya i have seen this many times in my cultures too,but not continuously. what i think is, this is this is not contamination, and is just a very thin n long particle that passes through filter sometimes while filtering media.what i suspect is this is a component of media powder what we get generally. think, when u have very good cleaned lab n clean work practice, where they should come from? ''MEDIA'' . i think u need not worry abt tat.

i am however little confused with disappearing of particles. are they dissolving slowly?

as i have seen in my medium, i think they are not moving, and they r not growing, ie., they r not contamination. if it is the case.dont worry. if not, take the medium containing this""hair particle"" n transfer to other plate, leave for few days, to see increase in size n number.

hope this would be useful.

Last edited by sweetsunil21; 05-17-2010 at 09:58 AM.
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