I'm testing cell viability on a novel platform we're building, and trying to see if the cells we're testing can persist without media when in a CO2 humidified incubator. Is CO2 + humidity enough for cells to survive for at most 2 hours? Or should I expect cell death when there is no media?
Currently testing HEK293 or L6 cells (immortalized), as well as a primary mouse embryonic fibroblast line.
Hello Doctor,
You mean cells will not be bathed in growth media? If yes, then you should expect cell death. Without growth media, the presence of CO2 and humidity will not serve any purpose.
CO2 (5%) is required to maintain the pH of the culture media containing the bicarbonate buffering system within the physiological range. Humidity is required to prevent evaporation of the culture media as evaporation may increase the concentration of salts in the media thereby having a negative impact on the cells.
Cells may survive without CO2 and humidity for 2 hours but not without culture media. You may culture cells without CO2 by adding about 15-20mM HEPES in the culture media without the presence of bicarbonate for nearly 8-10 hours depending on the concentration of the cells. HEPES help to maintain the pH of the culture media within the physiological range. Absence of humidity for 2 hours should not affect cell survival.
So, you should expect a decrease in cell viability even before 2 hours as CO2 and humidity are not going to help in cell survival.
Best.