Re: PBMC separation from brain tissue

From: Stojan Dimitrov (stoianski_fl@yahoo.com)
Date: Tue Apr 02 2002 - 09:52:31 EST


Dear Rick,
Why dont you try using the MACS system.
http://www.miltenyibiotec.com/index.php?site=products-human
Using anti-CD45 magnetic MicroBeads (for leucocytes)
you might separate all cells of hematopoietic origin
except erythrocytes, platelets and their precursor
cells.
Yours,
Stoyan

--- Rick Arthur Bright <rbright@emory.edu> wrote:
>
> I am trying to analyze lymphocytes, monocytes,
> granulocytes, etc from
> brain and lung tissues.  After collagenase digestion
> I, of course, have a
> lot of junk...stuff I don't want to run through the
> instrument.
>
> What is the best way of separating the cells from
> the junk?  I tried
> Ficoll-paque, which nets me the lymphocytes, but I
> lose other cells types
> that might be important.
>
> Would a Percoll gradient be a better route to go?
> What gradients would be
> most appropriate.  I am digging in the literature as
> well, finding many
> reference to percoll gradients, but few of them say
> what the gradients are
> and where the cells of interest will settle.
>
> Any quick thoughts would be helpful.  I, hopefully,
> will stumble upon that
> magic reference sometime tonight, but if you know
> already I would
> appreciate your help.
>
> Thanks
>
> Rick
>
>
>


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