Cultured cells at constant pH?
dspinella at chugaibio.com
Thu Aug 7 09:51:27 EST 1997
Ian York writes:
> This is probably a simple question, but I don't know where to start
> I want to grow tissue culture cells at a range of constant pHs for several
> days. The pH doesn't have to be fabulously accurate, but I don't want it
> to start dropping dramatically as the cells become denser. I'd like to
> try a range of, oh, I don't know, let's say 6.3 - 7.3 or so.
> Are there media that will do this? Where can I find recipes?
The problem with most culture media using the common CO2/bicarbonate
buffering system is that bicarb is a very weak buffer at physiological
pH. A better choice is one of the Good buffers such as HEPES (at 10 -
20 mM) which is a much stronger buffer at this pH range. See R. Ian
Freshney's "Culture of Animal Cells" for recipes (or just suplement your
standard media with 20 mM HEPES). Good luck.
More information about the Methods