Replies for Flow cytometric analysis of nonparenchymal cells isolated from liver tissue with collagenase

Jayaraman, Sundararajan anue2468 at uic.edu
Fri Nov 4 14:03:38 EST 2005


At least as far as the human islets are concerned, accutase is more
toxic than trypsin-EDTA. It kills more cells than trypsin-EDTA. One has
to be cautious about what is suitable for the specific cell type.

Sundararajan Jayaraman, Ph.D.
Associate Professor
Dept. of Surgery
University of Illinois College of Medicine
909 South Wolcott Avenue
Chicago, IL 60612
Phone: 312 355-5133 (Office); 312-413-0366 (Lab)
Fax:: 312-355-1497


-----Original Message-----
From: wang yilan [mailto:yilan1012 at yahoo.com.cn] 
Sent: Thursday, November 03, 2005 8:01 AM
To: cyto-inbox
Subject: Replies for Flow cytometric analysis of nonparenchymal cells
isolated from liver tissue with collagenase

Dear flowers,

Thank you for all your suggestions for my question.
Here I collect them as the following:

"William King" <wking at aecom.yu.edu>
Hello Yilan,
I would recommend a product called Accutase and for
tissues, AccuMax. Know that Accutase is much more
gentle than Trypsin. Accutase does not damage surface
antigens and no neutralization is necessary. Accutase
can be purchased from its creator, Innovative Cell
Technologies, or from all the usual suspects, e.g.,
Sigma, eBioscience, Chemicon, and probably many
others.
William


"Corver, W.E. (PATH)" <W.E.Corver at lumc.nl> 
Dear Yilan,
Use trypsin/EDTA or EDTA. Inexpensive and effective,
frequently without 
damaging cell surface epitopes:
Limited loss of nine tumor-associated surface
antigenic determinants 
after tryptic cell dissociation.
Cytometry. 1995 Mar 1;19(3):267-72. 
Best regards,
Willem Corver


"Jonni S. Moore" <moorej at mail.med.upenn.edu> 
we've had good luck with 5mM EDTA to remove the
adherent cells..just 
>add to flask, let sit for a bit at room temp (may
have to do a few 
>minutes at 37 if RT doesn't work. Immediately after
the cells come 
>off, transfer to cold EDTA containing staining buffer
and continue 
>with staining being sure to keep cold..this should
work for you..
jonni

"Ruth Bates" <r.l.bates at bham.ac.uk> 
I've found scraping works best with kidney
epithelials, people have 
published using just EDTA with the same cells as I use
but in my hands 
they won't come off even after an hour.
If your cells aren't too precious try EDTA first, then
scrape.

Ruth L Bates BSc(Hons) MSc PhD
Renal Research
Dept of Medicine
University of Birmingham
Queen Elizabeth Hospital
Edgbaston
B15 2TH
+44 (0)121 414 3785

http://www.medsciences.bham.ac.uk/staff/medicine/cockwell.htm


Yilan W


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