@saicharan - do we have any cell culture medium available with this HEPES buffer?
@ Chandrashekaram Kans- Hyclone make DMEM media is available with HEPES Buffer
Any one here can have comment on new designed incubator with Tri gases; nitrogen, CO2 and Oxygen plus controlled %RH? As far as I know, N2 gas used to control oxygen level in incubator, by doing this one can set his incubator to hypoxic condition reducing oxygen level to less than 5%.
I wonder if old fashion incubator having only CO2 monitor will be good enough for cell growth comparing with new designed which have %RH and oxygen level monitored?
In order to culture cells under optimum conditions the media they grow in needs to be buffered so it stays at neutral pH (around pH 7). It is possible to add a buffering compound to the media, like sodium phosphates or Tris or Hepes, but often these compounds affect the cells' growth. A carbonate buffer system, of HCO3- and H2CO3 has been found to affect the growth of cells the least. H2CO3 in water is in equilibrium with H2O+CO2. To keep the CO2 in the buffer solution at the right level you have to supply CO2 in the atmosphere above the buffer, otherwise all the CO2 would dissipate and the H2CO3 would disappear and there would be no more buffering effect (and the medium would become alkaline). So when you use a CO2 cell culture incubator, make sure the media contains bicarbonate buffer (usually from NaHCO3).
Khalid Omer Alfarouk
Indian Institute of Technology Bombay
University of Leipzig
Chandrasekaram Kans 430-2100
National Institute of Oceanography
Mahdi Habibpour Fatideh
Amirkabir University of Technology
City University of Hong Kong