I would be grateful if some of you could tell us about optimal CO2 and O2 conditions for human induced pluripotent stem cells (IPSC). We would like to cultivate these cells in pluripotent state, and also try to differentiate them into neurons.
Thansk in advance,
Ciao.D .
Like any other cells 5% co2 is sufficient to grow iPSC cells. Use the media recommended by iPSC. We grow the cells at 5% co2, 98% humidity, 37C nothing fancy. The cells are happy and they grow as expected.
Thanks a lot, Another question 5% CO2 in air, or do you balance levels of O2 by N2?
5% co2 in the incubator ( air). 95% air (humidity) takes care the rest. co2 helps to maintain the pH of the culture media.
You can read more about pH levels and C02 in this link
ttp://www.lifetechnologies.com/se/en/home/references/gibco-cell-culture-basics/cell-culture-environment/ph-co2-levels.html
pH Levels
Most normal mammalian cell lines grow well at pH 7.4, and there is very little variability among different cell strains. However, some transformed cell lines have been shown to grow better at slightly more acidic environments (pH 7.0 – 7.4), and some normal fibroblast cell lines prefer slightly more basic environments (pH 7.4 – 7.7). Insect cell lines such as Sf9 and Sf21 grow optimally at pH 6.2.
CO2 Levels
The growth medium controls the pH of the culture and buffers the cells in culture against changes in the pH. Usually, this buffering is achieved by including an organic (e.g., HEPES) or CO2-bicarbonate based buffer. Because the pH of the medium is dependent on the delicate balance of dissolved carbon dioxide (CO2) and bicarbonate (HCO3–), changes in the atmospheric CO2 can alter the pH of the medium. Therefore, it is necessary to use exogenous CO2 when using media buffered with a CO2-bicarbonate based buffer, especially if the cells are cultured in open dishes or transformed cell lines are cultured at high concentrations. While most researchers usually use 5 – 7% CO2 in air, 4 – 10% CO2 is common for most cell culture experiments. However, each medium has a recommended CO2 tension and bicarbonate concentration to achieve the correct pH and osmolality; refer to the media manufacturer’s instructions for more information.
Hello
i agree with Mohammed Hakim Jafferali , also see the attachment i hope help you .
Good Luck
Hi Davide,
As suggested already, standard tissue culture conditions (5% CO2 in a humidified incubator) work fine with iPS/ES, but you should minimize opening/closing the incubator, as these cells can be rather sensitive to environmental changes.
Hi All,
I will like to know if anyone know whether there is a difference between using 10 or 5% CO2 for cell culturing.
Ayodele Jacob Akinyemi The growth medium controls the pH of the culture and buffers the cells in culture against changes in the pH. Usually, this buffering is achieved by including an organic (e.g., HEPES) or CO2-bicarbonate based buffer. Because the pH of the medium is dependent on the delicate balance of dissolved carbon dioxide (CO2) and bicarbonate (HCO3–), changes in the atmospheric CO2 can alter the pH of the medium. Therefore, it is necessary to use exogenous CO2 when using media buffered with a CO2-bicarbonate based buffer, especially if the cells are cultured in open dishes or transformed cell lines are cultured at high concentrations. While most researchers usually use 5 – 7% CO2 in air, 4 – 10% CO2 is common for most cell culture experiments. However, each medium has a recommended CO2 tension and bicarbonate concentration to achieve the correct pH and osmolality; refer to the media manufacturer’s instructions for more information. Ref. https://www.thermofisher.com/za/en/home/references/gibco-cell-culture-basics/cell-culture-environment/ph-co2-levels.html
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