Cell culture being poisoined for nearly a year - Done by unknown person(s) (Aug/14/2008 )

I had posted several posts in BioForum (Cell Culture) asking people around why my cells died of no reason. I had been doing cell culture for 6 years, and since last autumn, my cells started to die (they are stuck to the bottom of the flask but very dead) and recently it got contaminated by the same bacteria. Only me in the lab has this problem (ie cells dying/getting contaminated continuosly). We all used the same cells, same media and shared incubators.

I thought I was the ONE which was doing things WRONG so I asked 3 person (2 technicians and my professor) to put cells in my incubator and write my name on the flasks (or no name). All the flasks which was with my name/no name on it end up dead or contaminated the next day but the original flasks (kept by the 3 other people) were all healthy and grew well.

So my professor sealed the culture flask with paraffin tape and sealed it in a bag (the cells are cultured in closed enviroment) but the cells he cultured died the next day. We found out the paraffin tape he sealed was TAMPERED (=resealed by someone else) cause he has a special method of sealing the tape. Other flasks which was contaminated has marks on the monolayer (cells got peeled of) due to harsh pipeting of liquid. Someone had added some liquid into the flasks after the cells formed a monolayer...

I then cultured the contaminated liquid and found that it was a strain of E. coli which was resistant to both ampicillin and kanamycin. Other cultures which were not contaminated but were dead looks like they were being killed by alcohol/some reagents to make it stick and dry on the bottom of the flask (I guess the motive is to fake 'natural' death)

I then shifted to another lab with restricted access (records whoever whom enters the lab, ID access) and ALL the cells I cultured were alive! And I used the same media which my cells used to die when I cultured them. No changes in methods, and I was able to do transfections and all was well.

I really do not know what to do. The lab with restricted access is only for temporary use and I certainly do not want someone to poison my cell culture again.

Why would anyone want to do such things? I had no 'special' relationship with anyone (I am only a foreign student in the research institute) and removing me from the institute does not benefit anyone (maybe only major humilation for my bosses if I quit)

Whatever it is, I am relieved because for nearly ONE whole year, I was not able to carry out any normal experiments and I kept on asking myself what wrong did I do. Even my another supervisor told me that my cells were dying of contamination because I wasnt good in aseptic techniques, etc. I hope, really hope that life can be much more normal from now on.......

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Wow, this is terrible, especially so that it has been going on for such a long time. I would think that your institution/PI would have to launch some kind of investigation and find the person? Certainly, you shouldn't feel forced to leave (and if you do, perhaps somebody else will be targeted). Maybe now that you have identified what the problem is, the person will know that you/the university will be looking for them and will now stop? But in any case, I wouldn't want to work with that person on any experiment - if they will do this in cell culture, would they change your reagents to water and other forms of sabotage around the lab?

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What does your professor plan to do? He must be really pissed off. There should be some kind of investigation like monitoring with CCTV or something of that sort.

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Thank you mRNAi man. My boss will talk to the head of the department. They wanted to keep things quiet for a while until the person is indentified. Meanwhile he wanted to put a camera near the incubator to investigate things.

Some of the lab members still think that I am wrong, even will all these evidence that me, my boss and the other technician had collected.

Anyway, the incident seems to happen early in the morning so I will get up at 5am and reach the lab by 6am (the lab opens at 6am) everyday till my project ends.

Cell death and contamination were recorded ONLY on the weekdays. I do cell culture almost everyday and if there are any contamination/cell death by accident/incubator problem, there is NO way that it happens to beautiful and healthy cells only on the weekdays and only happens in my cultures.

Please see
http://www.protocol-online.org/forums/inde...c=35872&hl=
http://www.protocol-online.org/forums/inde...c=32632&hl=

I am "lucky" that my boss is supportive now. He thought I wasnt doing my culture properly in the first place (that is why it took so long to find out what has happened).

I will fight this till the very end.

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I'm not sure whether going in at 6am is a good idea...to fire the person responsible your PI will need strong evidence. If you see somebody doing something to your plates, it would be your word against theirs. I think Bungalow Boy is correct - CCTV will be the best bet. In any case, I would keep your PI informed of what you plan to do - you wouldn't want to compromise any plan of action he wants to take. Or maybe he could go to lab early with you?

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There are small video cameras on sale, that look like a pen or something. Of course after talking with the professor. This person, whoever did that, should never do science again.

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Amazing and very sad indeed, they are in Science like in other jobs some really crazy people believe me I got to know two people like this One was trying to unplugged all the -80°C freezer of our lab as a revenge against my boss. The other one was even very dangerous she yell at the assistant director of my institute and tell him that she gonna kill him with a knife, she was arrested the next time she enter the institute after beeing fired

So you see that's sad but unfortunately not an isolated case

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If you don't mind me asking, how did you find the suspect? Any idea of her motive? Why is she doing this?

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Yes, he is planning to have a monitor.

He said he plans to fire the person soon. She is a post-doc with a medical degree (working as a doctor in a hospital too). Gives me chills to think about the patients that she does not like.

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We had records of who is entering the lab. Everyone has an ID. We determined the approximate time of cell death, and traced the person(s) who had entered the lab at that particular time. In such circumstances, someone which knows our lab layout very well, knows where we store our cells very well. And as I was the only target, knows where, how and when I store my things. And the person should also be very well aware of our schedules and should be able to enter P2 facilities without being suspected as an intruder.

So if we put all these pieces together, and find the time which the person entered the laboratory, things are actually quite simple from here. Besides, carrying antibiotics resistant E. coli to continously contaminate cells is not an easy job. I can have 25 T-flasks and multiple 24-wells plates, all running at the same time, so to contaminate ALL of them requires excellent knowledge of science and lab layout and habits of our lab people.

As for motives, I do not want to make assumptions, things can only be clear if you question the person but;

1. Jealousy
2. Pride, not getting enough praises from my boss. Overestimating one's worth
3. Hatred for my immediate boss (there were some misunderstandings between them)
4. Not getting punished even if caught, having confidence in never ever being exposed

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