cell dead after changing the medium to PBS buffer - (Apr/05/2007 )

Hello,
I encourted one problem in cultureing c2c12 cells.
I transfected the cells, and after 20hours I changing the mecium to PBS buffer, but it's so wierd that almost all cells dead in 30 minutes.
Is anyboby have some experiences about this phenomenon? Thanks.

-cynthiat6g-

You mean they came off the plate? They did so because there was no Ca and Mg in PBS, most likely.
How do you know for sure that they were dead?

-genehunter-1-

QUOTE (genehunter-1 @ Apr 5 2007, 02:38 PM)

You mean they came off the plate? They did so because there was no Ca and Mg in PBS, most likely.
How do you know for sure that they were dead?

Yes, the cells came off the plate and gatherd to clumps.
I thought the cells maybe dead.

Thanks

-cynthiat6g-

QUOTE (cynthiat6g @ Apr 5 2007, 11:46 PM)

QUOTE (genehunter-1 @ Apr 5 2007, 02:38 PM)

You mean they came off the plate? They did so because there was no Ca and Mg in PBS, most likely.
How do you know for sure that they were dead?

Yes, the cells came off the plate and gatherd to clumps.
I thought the cells maybe dead.

Thanks

phosphate has a similar effect as EDTA, and complexes Ca2+ and Mg2+ as genehunter-1 suggests; so cells are detached but likely not dead; such rapid destruction only would be achieved by harsher conditions; do a viability test or simply try to stain with trypan blue

-The Bearer-