Hello,

I would appreciate receiving comments about the detachment procedure for
cultured Mesenchymal stem/stromal cells (MSCs) for FACS analysis.

I am starting immunophenotyping these cells and I am concerned about
damaging surface antigens using trypsin; however, trypsin is the only
procedure that is working in my hands right now:

other methods I tried:

EDTA and a Cell dissociation buffer (Enzime free, PBS based) from Gibco;
these last two methods were not succesful since the cells formed very
tight clumps which did not come out with filtering methods, and lots of
the cells died.

So I am using trypsin at a low concentration ( 0.05%%), and let the cells
stand in medium with serum for one hour to let them recover from the
trypsinization process.

Do you have a detailed procedure for this, with particular emphasis on
(1) trypsin-EDTA concentration (2) time left between trypsinization and
recovery of any damaged surface antigen until FACS analysis.

Any thoughts are welcomed

Thank you in advance.

Hi,

Do both extracts, water and alcoholic.

lit

--- On Sun, 29/6/08, immuno-request@oat.bio.indiana.edu oat.bio.indiana.edu> wrote:
From: immuno-request@oat.bio.indiana.edu oat.bio.indiana.edu>
Subject: Immuno Digest, Vol 34, Issue 2
To: immuno@magpie.bio.indiana.edu
Date: Sunday, 29 June, 2008, 1:05 AM

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