I'm growing MCF7 cells and I was told that in order to avoid contamination it is best to have a separate small room that is sterile with the hood and everything else that is necessary for culturing cells. I was just wondering if this is normal or if it is possible to keep a healthy cell line in a large open laboratory without contamination. Thank you!!
I tend to agree with above, you do not need a separate room but it is ideal. It is probably better to use a room that does not have a lot of people moving in and out doors opening affect the flow of air through a hood. It is always a good idea to clean hoods before and after use and to not leave anything in them between users. Depending on the hood in use a thorough clean before starting new cell lines can be a good idea class II hoods can get quite dirty if media is spilt through through the front grill and removing the bottom and cleaning underneath is a useful house keeping job. Windows should not be open in a room with a hood, air conditioning is better but ensure this does not affect the hood's airflow again. If you are starting a new piece of work in a hood not used before fumigation should be considered as the starting point. All your materials should be your own to ensure sterility this includes media and tissue culture plastics (disposable) if using shared equipment such as pipettes these should be cleaned before use wiping with 70% IMS is one way.
A separate tissue culture facility is the ideal situation and may be essential depending on the nature of the experiments and the cell type you plan to use. Contamination with mycoplasma is always an issue and I would recommend you check for this at regular intervals.
I agree with the above. Clearly, in an academic setting, your colleagues sharing the LAF bench could impose a bigger problem than the fact that the bench is placed in a regular laboratory. You should implement cleaning lists and other routines such as consensus aseptic rules etc to minimise risks for cross contamination. Have the bench regularly serviced and keep the surrounding as clean and free of dirty materials as possible. The same should go for shared incubators of course...
I tend to agree with above, you do not need a separate room but it is ideal. It is probably better to use a room that does not have a lot of people moving in and out doors opening affect the flow of air through a hood. It is always a good idea to clean hoods before and after use and to not leave anything in them between users. Depending on the hood in use a thorough clean before starting new cell lines can be a good idea class II hoods can get quite dirty if media is spilt through through the front grill and removing the bottom and cleaning underneath is a useful house keeping job. Windows should not be open in a room with a hood, air conditioning is better but ensure this does not affect the hood's airflow again. If you are starting a new piece of work in a hood not used before fumigation should be considered as the starting point. All your materials should be your own to ensure sterility this includes media and tissue culture plastics (disposable) if using shared equipment such as pipettes these should be cleaned before use wiping with 70% IMS is one way.
I also agree with everyone's fact/opinion. The most important thing also, you should avoid crowding the incubator with lots of different cell line, as it may be the source of contamination since there is no sterility control (air flow) inside it. Also you need to ensure that the water bath (if you are using one to warm up your media etc) is properly clean , I wash mine with disinfectant bi-monthly since I'm the only user.
Well, I think is not strictly necesary but is a good protocol and idea to avoid contamination. In my lab cells are growing in differents rooms.
I do suggest to perform cell culture work with cell lines and primary cells in separate labs
Under my point of view, everyone is stating very good points. Personally, I think that if you are just growing MCF7 cells, you could get away with not using a separate room providing you are following all the wise suggestions people above have mentioned. Just make a good frozen working bank for two reasons. One, avoid too many passages of cells (otherwise they will eventually become differentiated somehow) and two, in the case you have a clear contamination (normally fungi or bacteria), you can always start all over again with a fresh frozen vial from your working bank. Finally, do not forget to check regularly for mycoplasma contamination. This contamination will not be obvious through the microscope!
We just set up a human tissue culture lab few months ago, as it is required by the university within the occupational heath and safety framework. We need to have a PC 2 lab and a bio safety cabinet. All the cell culture technique requires sterile handling and the protocols for PC2 lab differs to PC1 lab. In this environment, the cells line can grow without contamination, also it prevents us from any contamination. I know that cells will not jump at us and contaminate.
Referring to your question now: it is compulsory to have a lab dedicated to tissue culture. I am not sure to grow cells otherwise.
Hope this helps
The bio-safety cabinet (hood) and good tissue culture techniques (some of which are listed above) keep the cells free of contamination. A separate room helps by minimizing traffic, keeping laminar air flow in the hood when in use. A minor effect is that the running hood also HEPA filters the air (but this is not as important and using the HEPA filters like that is pricey).
Separate rooms for Class 2 tissue culture are not so much a principle of maintaining clean cell cultures, but of maintaining a clean laboratory and protecting people who work in the lab. MCF7 cells, being human cancer cells, can carry human pathogens and potentially cause tumors in human. By having a separate room and keeping the living cells in it, it ensure that the rest of the lab is safe. Also, if there is a problem, it is easier to section off and clean the designated culture room, than the whole lab. Good practice is that cells should only be removed from this room if the cells are dead (lysed, fixed, bleached, etc.). This kind of stuff should be institutional policy and what safety officers look for.
If there is another lab/lab members that have a Class 2 lab setup, you should be able to use theirs. Just keep things clean and sterilize common equipment between uses. I worked in a lab with HeLa and primary mouse lines in one incubator, let alone room. MCF7 do not need a separate room from other Class 2 cells and up. Someone may want to keep these types of cells out for their work area for personal safety or because cancer cells like MCF7 or HeLa more easily cause cross-contamination to their cells, but this is personal opinion.
It's part of the good laboratory practice to separate cell culture into an enclosed space. However, a lot contamination depends on a lot of factors. I have good reason to believe that water baths are the number 1 contamination source. More than bacterial contamination culture cross-contamination is the biggest threat. It is highly suggested that labs use separate hoods for handling transformed cell lines and primary cells.
Dedicated facilities for culture of each cell type is not needed and very costly at Research stage.But one need to have a very good cleaning validation programs and records ( one can have two or three laminar flow hoods in same cabins at R&D stage)
But if the tissue culture work is at larger and regular commercial purpose it is essential to have a dedicated facility.
As others have indicated, a separate room is ideal, but not necessary for MCF7 unless it is mandated by your institution. To my knowledge MCF7 does not harbour any infectious agents when obtained from reliable sources like ATCC, although this should be confirmed for your specific cell line, so it's use requires only the care that any human cell line would. Remember that some of the most important bacteriology was performed for more than a century on open benchtops in open labs and early mammalian cell culture was done for decades on open benchtops without laminar flow or even still hoods. Good sterile technique, general lab cleanliness, and common sense go a very long way toward success in cell culture. That said, a separate room, a laminar flow hood, and tightly limiting access to all tissue culture supplies (i.e., everybody has their own bottle[s] of media...not shared) also help a lot.
it is good if you have a separate room for cell culture , because it gets contamination if you have so many people around culture room .
If you are a beginner in cell culturing it is better for you to have a separate room for tissue and cell cultures. This will make it easier to you to maintain sterile conditions and reduce the contamination of cells. You will be less distracted and the whole process of cell culturing will be easier to control. All the necessary culture supplies for the work with cells will be on hand.
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