We are attempting to culture various cell time at narrow ph margins. Ph of media is optimized to ranges of -+ 0.05. This can be attained at a constant level for several weeks at a time however we run into occasional drastic ph changes of -+ 0.1-0.2 on an irregular basis. We calibrate ph meters daily at 3 points and measure Co2 concentration of incubators daily with infrared meter. Mycoplasma antibiotic is added to our media and has been confirmed negative to infection via PCR. Our media formulation is consistent so unlikely due to changes in media stocks. Any feed back would be appreciated.
Do you culture the cells in batch cultures (Flasks, petri dishes, etc.) ? Do you consider the metabolism of the cells. It will affect te pH. Or do you use culture vessels with permanent flow of medium coming from a large medium reservoir?
Hi Matthias,
these are embryo cultures under oil. There is a only 1-3 embryo's per 200 micoL drops. These we run many of embryos using this technique and never have a problem with ph changes for weeks on end. Its only occasionally we run into the ph changing. When deviates from the norm its almost always shifting to alkaline.
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