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Keratinocytes Chemotaxis Assay


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#1 NazGoole

NazGoole

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Posted 10 April 2014 - 11:09 AM

Hi,

 

After some testing (and failing), I am soon doing some "serious" chemotaxis assay with keratinocytes. I will be using uncoated 8 micron FluoroBlok filters and staining the cells with calcein. The upper chamber will be filled with unsupplemented serum-free kera growth medium (with 1% BSA), and the bottom well with adipose tissue conditioned medium.
One problem I have is that I dont know "which" keratinocytes to use best. As keratinocytes are present in different stadiums of differentiation. Should I use "early" highly proliferative ones? Or rather older broader and more differentiated ones? How long do you think I should incubate the system? My supervisor estimated to wait around 2-5 hours until measuring. Whereas some papers reported that migration started only after around 24 hours....
The upper chamber will be filled with unsupplemented serum-free kera growth medium (with 1% BSA), and the bottom well with adipose tissue conditioned medium.

 

Thanks a lot!






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