I got my HEPG2 cells passaged to a new cell culture flask and have mistakenly used 10 X PBS for washing instead of 1 X PBS. Although washing took relatively short time how bad might be the consequences? Are there any experts in cell culture who could share their experience. I thank you in advance!
It is my understanding PBS is a wash buffer, but in strong concentrations may lyse cells as it acts as a detergent. If your cells are confluent and look healthy, I would not worry about this mistake. I would suggest you look into switching to HEPES-buffered saline instead of PBS. I also used PBS until my new mentor told me HEPES is much more gentle on cells compared to PBS. I also found it was cheaper to make than PBS. Good luck with your culture.
Hi Stefan,
That happened to one of our undergraduate students.
There is no point wasting time on saving cells that might have been stressed/lysed if you have frozen vials of your cells.
Good luck
Hi Stefan,
if you used 10x PBS in your cells you added a solution with 10 times the osmolarity of a physiological solution, which would produce your cells to shrink produced by the flow of water form the cytosol of the cell to the PBS. Once you put the cells in the regular media the correct osmolarity would make the water to reintroduce the cell, however the cells might be already too damaged. You could keep the cells and see how they look the following day but I would recomend you to just toss them and start with new cells.
Hope this helps.
Ezequiel
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