Hi to all,
I need to incubate a plate of hela cells in a solution of PBS (without Mg and Ca). Is it dangerous? do PBS cause detachment or morphology changes of the cells if I incubate them for 30 min in this buffer at 37 °C?
thank you
HeLa cells are gerenally quiete robust and the incubation with PBS is not dangerous.
Although i have to say that i never incubated them in PBS at 37°C for 30 mins, I think it might result in detachement.
I freequently use !cold! PBS to detach adherent tumor cells like HEK and others.
regards
I am not sure what your application is exactly. I routinely wash and have them in PBS at RT for 30 min-1 h for cell scans. They will begin to round up after 30 min and start detaching after 45 min or so. I don't have any downstream issues even after an hour when harvesting for protein/RNA analyses. I did leave some out for >2-3 hours but won't know the answer until a few weeks from now. I have not tried to add Mg+/Ca++ to help them stay attached but have added 7% FBS-PBS which helped maintain morphology/attachment over 30-45 min, if i recall correctly. Hope this helps!
In PBS 15 minutes to 30 minutes the cells will remain alive. If you want to keep for more time better you use phenol free HBSS buffer. Up to 1 hour you can keep your cells intact in this particular buffer.