Hi
I would like to ask if washing cultured adherent cells with PBS is unmet befor starving cells. In this case, how we can avoid loosing cells during washing steps. Does the rate of disacared cells from wells will be approximately the same?
Thanks
For testing growth in a serum-free media you need to wash at least twice in PBS to avoid any carryover effects. For general purposes, one wash is enough.
"Regular" adherent cell lines do not detach at all while washing with PBS. If you use less adherent cells like HEK293 then you can wash them with a Ca/Mg-containing PBS (absence of these metals decreases adherence) or grow them on poly-lysine treated plates (provides a strong adherent for HEK).
it is very necessary to wash your before serum starvation step.
I recommend that adherent cells should be washed gently before serum starvation step.
good luck
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