Hello all,
We want to scale up cell culture facilities in our lab from flask to roller bottle. Usually we use CO2 incubator for cell culturing activity in cell culture flask. My question is...how important CO2 is in cell culturing activity using roller bottles? The cell line is VeRo and will be used for virus production.
Thank you.
Dear Khrisna,
the CO2, cells are nomally cultured in, is important for the buffer system of the medium. As far as I know, cells in Roller bottles are also cultivated in a 37°C/5% CO2/humid incubator. But in case you don't want to or you cannot afford buying a special incubator, there are also special rollers available that are fitting in a normal incubator.
I hope this information is helpful for you!
Hello Khrisna,
The media used for culturing mammalian cells have Sodium-bicarbonate/have to be supplemented with the same, as it acts as a buffer. You can normally grow vero cells in cell culture flasks as well as rollers (Closed system) for virus production, without the need of 5% CO2 atmosphere, whereas for cell culture plates (96 well, 48well/any plate, which all are open system) we need 5% C02 atmosphere. The only difference in the media used for closed and open system is the concentration of Sodium-bicarbonate. The CO2 media has higher concentration of Sodium bicarbonate than non CO2 media. The rollers and flasks needed to grow vero cells without 5% CO2 atmosphere should not have vented caps for gaseous exchange, and you can use non CO2 media having low concentration of Sodium-bucarbonate without 5% CO2 atmosphere.
Hope this information will help you
Dear Ariane,
Thank you for your answer. Do you use different type of media like Dheeraj mentioned below your answer? Or it's just 100% same with cell culture in flask?How about in virus inoculation process? Will CO2 availability alter the process?
Dear Dheeraj,
Thank you for your answer. So if we want to use CO2/not it really depends on media type, isn't it? How about in virus inoculation process? Will CO2 availability alter the process?
____
Still open for other opinions...
Dear Khrisna,
there are different possibilities:
1. You can use the normal medium which is containing the CO2-dependent buffer system and you use Roller bottles with vented caps (gas exchange has to be possible)
2. You can use the normal medium and you use Roller bottles with normal closed caps, then you have to make sure that there is enough headspace in the bottles (don't fill them completely up) and you have to sparge the medium with CO2 before closing. Then you have to check very often the status of your medium/cell suspension
3. You can use special CO2-independent medium, but you have to check if it's available for your specific cell line.
Normally you cultivate cells in Roller bottles with CO2 and I don't know if it's important for your special application, but I don't think that the CO2 should have an impact on this if you are using the right medium, since it's only required for the buffer system!
Hello Krishna,
The media type i discussed about: CO2 Medium and Non CO2 Medium are our Labs's classification, they are not commercial.
CO2 medium has higher amount of Sodium bicarbonate and NonCO2 medium has lower amount of sodium bicarbonate. CO2 and Sodium bicrbonate present in culture medium make a buffer system.
So, technically if you need to switch from CO2 dependent culture to CO2 independent culture, you need to reduce the amount of Sodium Bicarbonate in your Culture medium.
Now,
If you are using ready to use liquid medium, you cant do so.
In our lab we culture VERO cells and produce Virus. For culture medium we use EMEM powder (Sigma-aldrich), and we need to add Sodium Bicarbonate and L-Glutamine while preparing the media. So we have the flexibility to prepare medium for a CO2 dependent/CO2 independent cell culture system.
You can follow the link :
http://www4.mpbio.com/ecom/docs/proddata.nsf/5f64ffd4f38c2fda8525645d00769d68/d129d700747af023852568b200619fff
Hope this information will help you
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