Re: Protocoll for mouse DC

From: Adrian Smith (A.Smith@centenary.usyd.edu.AU)
Date: Fri Feb 23 2001 - 18:29:48 EST


At 1:53 PM +0100 21/2/01, Kerstin wrote:
>Hello everybody,
>does anybody know a protocoll/paper for isolating DC of Mouse, label them
>with CFSE, bring them back in a mouse and search later for their
>localization?
>Thanks Kerstin
>


We have done this with CMFDA and it was published in JExMed in 1999.
The reference is

Smith, A. L. and B. Fazekas de St. Groth (1999). "Antigen-pulsed
CD8a+ Dendritic Cells Generate an Immune Response after Subcutaneous
Injection without Homing to the Draining Lymph Nodes." Journal of
Experimental Medicine 189(3): 593-598.

I have done it with CFSE but it can be tricky because CFSE can be
quite toxic to the cells than CMFDA so you have to buffer the
labelling solution with extra protein (the paper is only a Brief
Definitive Report so the methods are fairly abbreviated).

Feel free to ask any specific questions.

Hope that helps,

Adrian

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Adrian Smith			 T CELL BIOLOGY GROUP
Centenary Institute of Cancer Medicine & Cell Biology
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