From linalit from yahoo.com  Tue Jul  1 09:51:50 2008
From: linalit from yahoo.com (lit lei cheng)
Date: Tue Jul  1 11:28:29 2008
Subject: [Immunology] Re: Immuno Digest, Vol 34, Issue 2
In-Reply-To: <200806281705.m5SH57O27163@net.bio.net>
Message-ID: <798735.6096.qm@web32404.mail.mud.yahoo.com>

Hi,

Do both extracts, water and alcoholic. 

lit

--- On Sun, 29/6/08, immuno-request@oat.bio.indiana.edu <immuno-request@oat.bio.indiana.edu> wrote:
From: immuno-request@oat.bio.indiana.edu <immuno-request@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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Today's Topics:

   1. NEED GUIDANCE (ashis rasaily)


----------------------------------------------------------------------

Message: 1
Date: Fri, 27 Jun 2008 10:38:51 +0530
From: "ashis rasaily" <ashis.rasaily@gmail.com>
Subject: [Immunology] NEED GUIDANCE
To: immuno@magpie.bio.indiana.edu
Message-ID:
	<e2a5c43c0806262208l4ea08421hd258472ea47fcc48@mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

   I HAVE A FUNDAMENTAL QUERY REGARDING MY PROJECT [ immunomodulatory
  action of aloe vera and andrographis paniculata in human pbmcs].
  a] what extract[ alcoholic or aqueous] of the above mentioned plant
  extracts should i use??
  awaiting your reply.......................ASHIS



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End of Immuno Digest, Vol 34, Issue 2
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From MBolontrade from leloir.org.ar  Wed Jul  2 12:24:39 2008
From: MBolontrade from leloir.org.ar (Marcela Fabiana Bolontrade)
Date: Wed Jul  2 12:55:39 2008
Subject: [Immunology] Immunophenotyping MSCs
Message-ID: <444FCD3FB82E544C8565522DE4D0A49D03764123@munich.leloir.org.ar>

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.

 

Marcela