We have to optimize the growth characteristic for cell line culture. Ph is a very important factor for this. We want to use both RPMI and DMEM medium in the same CO2 incubator. But the CO2 requirement for these two different media are different. How can we manage this?
Bivek,
You can try using less Sodium Bicarbonate (Use powdered formulation of DMEM) and supplement with half the amount of Sodium Bicarbonate (Tissue Culture grade) than used in your medium which recommends 10% CO2.
I have used DMEM using 5% CO2 for years and have never had a problem. Just give it a try on a test sample. If the test fails add HEPES buffer as stated above. If you use HEPES buffer make sure to test your cells because some cells are sensitive to even low concentrations of HEPES
It will depend on the cell density in the DMEM. At low cell densities the pH may be higher than in RPMI
Thank you all for such valuable information. I cultured Huh-7 and LX-2 cell lines. Lx-2 cells have grown faster than Huh-7. There is more confluency in LX-2 after 24 hours of incubation. So I concluded that that DMEM can be used at 5%CO2 but its buffering capacity is at 10%CO2. So we can use same incubator for both DMEM and RPMI and HEPES is also toxic to some cells.
http://www.emdmillipore.com/US/en/product/LX-2-Human-Hepatic-Stellate-Cell-Line-,MM_NF-SCC064
- Badges
- Science method