Splitting cells: Why It’s Important and 3 Essential Tips | CST Tech Tips

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Splitting cell lines is an essential lab skill. This video covers the background of why regularly splitting cells is important for maintaining healthy cell cultures. We will also show you how splitting ratios work, and highlight three essential tips for subculturing mammalian cell lines.
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[@funny11744]

What is the value of humidity in the CO2 incubator?

[@cellsignaldotcom]

Hi, this varies depending on the cell line. Please refer to information from the cell line supplier on optimal growth conditions for your specific cell line. For mammalian cell lines, 5% is the most common CO2 concentration, but some cell lines may require 7.5% or a different CO2 concentration to properly maintain pH of the media, which is typically buffered with a bicarbonate-based system.
By the way, if you are culturing insect cell lines, these will have a lower temperature (again, check with the supplier) and likely do not require maintaining a specific CO2% since the media used for these cell lines isn't buffered with bicarbonate.

[@funny11744]

@@cellsignaldotcom THanks. I meant the value of humidity not the value of CO2 .

[@cellsignaldotcom]

Oh sorry about that! Most incubators don't actively adjust humidity levels the way they do for C02; many users place a pan of water in the bottom of the incubator and include a bacterial/fungal inhibitor. This is sufficient to reduce the rate of vapor evaporation from cell culture vessels, as long as they are being consistently maintained and checked.

[@funny11744]

@@cellsignaldotcom many thanks

[@kosheeka]

@@funny11744 A high humidity is recommended (around 95%) to minimize water evaporation from cell culture media. Evaporation can lead to changes in concentration of salts and other components, harming the cells. Opening the incubator door can cause a temporary dip in humidity due to cooler room air entering. This is why it's important to minimize the frequency and duration of door openings.