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Problem with transwell migration

transwell migration

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4 replies to this topic

#1 yi198720022004

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Posted 12 February 2012 - 06:43 PM

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!

#2 Inmost sun

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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 scolix

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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 yi198720022004

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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 wernerlael

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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?






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