Is it Okay to provide 5%CO2 to insect cells SF21? - (May/01/2008 )

Hi guys,

I am trying to culture SF21 cells in our lab,. Now there is a CO2 incubator available, which is used to be a mammalian cells incubator. I know CO2 is not required for insect cells. The problem is that I couldn't disable the CO2 supply. Anybody has any experience? Thanks very much.

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You might also have trouble with the temp. Mammalian cell lines (all I know of) are grown at 37degrees. These cells have to be grown around 28degrees and will die at temps above 30. As for the CO2, there are culture flasks that have plug caps and don't allow for gas exchange. You can grow these cells in ambient temps on a benchtop or in a drawer, they will just grow a bit slower than if kept at 28degree. Check out this manual from Invitrogen:

www.tools.invitrogen.com/content/sfs/manuals/insect_man.pdf

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Ideally you need a cooled incubator....we use an LMS cooled incubator. We however grow our cells at 21 oC and INFECT at 28oC. In our experience if you grow the cells at 28 they grow far to quickly.
Getting back to the question, as stated Sf9/21 do need need CO2. Just set your incubator to 0% CO2......or USE CLOSED TISSUE CULTURE FLASKS and NOT use ones with filtered caps. We grow our cells in Techne Stirrer bottles....far easier and more convenient. You can put non vented caps on these bottles.
HOWEVER a standard CO2 incubator will STRUGGLE to maintain a 28oC temperature, as they are affected by ambient temperatures.

Kindest regards

Rhombus

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Thanks so much for your reply.We don't have the temperature problem. Our incubator maintains at 27oC just well but I couldn't set to 0% CO2 because it would keep beeping. Now I am using 1% CO2, hope my cells are fine.

Here comes another question--what is CO2 for in mamalian cell culturing? to maintain the pH? or?
Anybody has an idea?

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CO2 is to maintain pH if you use a carbonate buffer, which is the case with the mammalian cell culture medium.
It will acidifie the medium.

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