Problem with transwell migration - (Feb/12/2012 )

Hello,

I am new in doing migration assay. I have problem maintain the cell viability after 3 hr of migration. I only get 8% to 12% cell that are alive after migration. I used 0.5 BSA migration medium and 3% FBS migration medium and FBS give me just a little better viability. I just don't understand how they all die after just 3hr of migration. Can someone give me some tips on migration assay please?

Thanks in advance!

I suppose you work with adherent cells, you should over coated wells; which FBS % do you use in normal cell culture?

some cells are not suitable for migration. Has this cell line been described in literature for these experiments.

I am working with black 6 mouse splenoytes. I did total splenocytes migration and I am looking specifically for Th17 subset. I used 10%FBS for normal cell culture. I read it somewhere says migration assay should avoid FBS becuase the cell will migration nonspecifically that's why I used BSA but they died.


How do you know they are dead?
Did you check if they are still ABOVE the chamber?
perhaps they are still clumps and can"t pass through Transwell?