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!
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Problem with transwell migration
Started by yi198720022004, Feb 12 2012 06:43 PM
transwell migration
4 replies to this topic
#1
Posted 12 February 2012 - 06:43 PM
#2
Posted 17 February 2012 - 06:57 AM
I suppose you work with adherent cells, you should over coated wells; which FBS % do you use in normal cell culture?
#3
Posted 17 February 2012 - 09:59 AM
some cells are not suitable for migration. Has this cell line been described in literature for these experiments.
#4
Posted 17 February 2012 - 09:57 PM
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.
#5
Posted 16 October 2012 - 01:11 AM
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?