In our cell culture lab, the incubator has a problem with making CO2, and all our cells are in there. Do you have any idea to make CO2 in the incubator or any idea about protecting our cells while we fix our incubator.
You can make CO2 from sodium carbonate/bicarbonate and a strong, non-volatile acid were the salt is reasonably water-soluble. Be aware that the chemical reaction produces heat and large volumes of gas!
Hello Nafiseh Paysokhan
Yes, it is possible. You take an empty jar made of glass of a suitable size that has a cover as well as candles. You need to sterilize the jar, cover, and the candles. Place the lighted candles and the cell culture flasks inside the glass jar. Close the jar with the cover with the candles burning inside the jar. Make the glass jar airtight by sealing the jar to avoid any exchange of gases. If the candles inside the glass jar goes off it means that the CO2 concentration inside the glass jar is around 4.5-5%. However, this method would be painstaking.
Alternatively, you could use the culture media containing HEPES at a concentration between 10mM and 20mM. Unlike the bicarbonate buffers which require CO2, HEPES containing media do not need CO2 atmosphere for buffering. When HEPES is present in culture media, the use of bicarbonate should be avoided. But if the cell density is low, absence of bicarbonate may limit the growth of cells. In case if bicarbonate is also present in the media, the concentration of HEPES must be about double that of bicarbonate to have maximum buffering capacity.
HEPES buffer can generate toxic reactive oxygen species in combination with tryptophan and other media components if the media is not protected from light. So, if you decide to add HEPES in the culture media, keep it stored in bottle covered in aluminium foil to protect from light. Perform media handling in darkness as much as possible to prevent this effect. Please note that the use of such media containing HEPES is viable for no more than 10-12 hours and is highly dependent upon type of cell line and cell concentration.
Best.
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