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-   -   Splenic adherent cells wont adhere! (http://www.molecularstation.com/forum/immunology-host-pathogen-interactions/71056-splenic-adherent-cells-wont-adhere.html)

yzfmonster 10-30-2009 02:22 PM

Splenic adherent cells wont adhere!
 
Hi, all
I am newbie to immunology and trying to culture mouse spleen adherent cells to study cytokine production upon LPS stimulation.

I dissect spleen, mechanically dissociate using the end of plunger, and plate onto petridish/culture dish in RPMI1640/Heat Inactivated 10% New born Calf Serum/Pen/Strep for one hour.

After one hour incubation, I can see lots of cells adhere. :happy: So I wash 3 times to remove non-adherent cells and incubate overnight. Then for some reason almost all cells come off the plate and not much left on the plate!
:pissed:

Please Immune expert here could you provide any solution/mistake I am making?
Thank you veery much in advance!

yzf


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