Does anyone else gas their cells like this?! - (Jan/09/2009 )
Hi,
I have recently started in a new lab where I am growing suspension cells that have to be maintained in the usual way - in an environment with 5% Co2,at 37oC etc. However, I have been told that I have to manually gas my cells with 5% Co2/air instead of growing them in a CO2 incubator. So when I passage my cells I add cell suspension, then medium, then I have to add CO2 from a gas cylinder into the flask, before tightly shutting the flask lid and putting my cells in a normal (non Co2) 37oC incubator.
Does anyone else do this?? 
I have 3 years cell culture experience and have never come accross this way of growing cells. I'm just a bit worried about it!
There are even CO2 incubators in the tissue culture room I use that aren't being used!
Someone please tell me my cells will be ok!! 
Thanks!
I never heard anyone put any gas directory into culture flask.
just out of curiosity, what is the purpose of that??
When we grow our cells in roller bottles, the rolling apparatus is in the non-CO2 37C incubator, so we do as you say and add CO2/air to the media. Your cells will be fine! But if you really can't get used to it, why not ask if you can use one of the CO2 incubators for your work??
I have recently started in a new lab where I am growing suspension cells that have to be maintained in the usual way - in an environment with 5% Co2,at 37oC etc. However, I have been told that I have to manually gas my cells with 5% Co2/air instead of growing them in a CO2 incubator. So when I passage my cells I add cell suspension, then medium, then I have to add CO2 from a gas cylinder into the flask, before tightly shutting the flask lid and putting my cells in a normal (non Co2) 37oC incubator.
Does anyone else do this??
I have 3 years cell culture experience and have never come accross this way of growing cells. I'm just a bit worried about it!
There are even CO2 incubators in the tissue culture room I use that aren't being used!
Someone please tell me my cells will be ok!!
Thanks!
Dear Confuzzled,
In 30 years of Tissue culture I have never heard of this.
Question: What cells are you growing?
If they require 5%CO2 then use a CO2 Incubator. Manufacturers have developed IR CO2 Sensors that are very sensitive and keep the incubator's stable at 5%.
We do use non CO2 Incubators for cell culture, but this requires using HEPES buffered media in closed TC flasks.
Kindest regards
Rhombus
I was once told that tissue culture flasks only need to be in CO2 for the first few hours after passaging as after the cells adhere they will start respiring and produce their own CO2. I guess with suspension cultures they are always actively respiring and produce enough CO2 to maintain the pH of the media???
Dear confuzzled,
don't worry, we have done this for years and years and years ... Our department is quite big, and purchasing that many CO2 incubators would be a big bite out of our budget. We have CO2 incubators, but these are reserved for plates that cannot be tightly closed. When using tissue culture flasks, we always introduce 5% CO2 gas mixtures before closing them. We have two large 'hot' rooms at 37°C where we can put these flasks. This is beneficial for two reasons: the CO2 incubators are opened a lot, and as such, the temperature, humidity and CO2 levels are not constant (so our plates do 'suffer' from this, but at least our flasks remain at a constant temp/humidity/CO2 level) and secondly, some people have observed less contaminations when using closed flasks instead of leaving the flasks open in the CO2 incubator (which we are actually not allowed to do).
@Rhombus, I know, leaving flasks open in a CO2 incubator is normally not a problem, but the high turnover of people in our department results in a lot of improperly trained/clumsy people (I know, not a good excuse, but it's reality, and to avoid those people messing up my experiments, I'm willing to do the extra step)