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What is the intracellular pH in cultured cells

Raymond Chang raymond.chang at icf.med.uni-muenchen.de
Fri Jul 21 13:41:39 EST 1995


To all cell culture scientists,

	Have you tested the intracellular pH of your cells? We have used
BCECF fluorescent dye to test the intracellular pH of C6 glioma cells for
quite a long time. We often have a pH of about 7.15 - 7.3. However, the
intracellular pH of the cells suddenly going up to 7.4 - 7.45 even the
extracellular pH keeping on 7.4 in the last two weeks. We have three times
such experience before. I found that perhaps the cell density was too high
because of the fast growth rate of C6 glioma cells. Then I subcultured
them in a lower density and waited for several days, the pH went back to
our normal record (i.e. 7.15 - 7.3). If I follow this experience, the
reason behind will be too high cell density give high pH. However, this
will be the opposit phenomenon as we generally accepted (i.e. cell
spreading leads to high intracellular pH) because the cells are round in a
high density. Is there anyone has any idea of this phenonmenon 

	Someone in the lab think perhaps there is mycoplasma
contamination. We have tested this before and found no mycoplasma although
we have not tested this time. However, my question is will mycoplasma
contamination leading to the change of intracellular pH? If so, will they
shift to alkaline or acidic? My personal opinion is I do not think so but
I cannot give a good explanation of this phenmenon. Can anyone give me the
suggestion. Thank you in advance.

Raymond, C.C. Chang,
DAAD Research Fellow,
Laboratory of Neurobiology,
Institute for Surgical Research,
Klinikum Grosshadern,
University of Munich,
Munich, 
Germany.





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