RE: Single cell murine skin suspension

Dear all,

Following this question, I would like to know where to find a detailled protocol to
harvest cells from murine skin. 

Many thanks in advance for your help.

Karine

-----Original Message-----
From: Sydney Lu [mailto:lus@mskcc.org]
Sent: 23 January 2006 20:58
To: cyto-inbox
Subject: Re: Single cell murine skin suspension


If you're interested in CD45+ cells from murine ear epithelium, expect 
at most 1 million cells from a pair of ears. Of these, 50% are 
gammadelta-T cells and 50% are CD11c+. I've tried to use skin from the 
body, but with no real luck.
In my hands the enzymatic digestion steps necessary to get to a 
suspension also kill quite a few cells, but I'm not sure if there's a 
good way around that.



Dr. Tomas Corcoran wrote:
> 
> 
> Greetings all,
> 
>  
> 
> A query- has anyone made single cell suspensions from Murine skin ? We 
> seem to be losing a lot of numbers with primary and secondary antibody 
> staining. Not sure what we are doing wrong although I suspect that the 
> repeated washings are resulting in exponential losses. Should we be 
> using a single flurochrome-labelled step ? Any help would be greatly 
> appreciated.
> 
>  
> 
>  
> 
> Tomas Corcoran
>