coagulation in leukapheresis

Durett, April G. agdurett at txccc.org
Sat Sep 30 00:28:44 EST 2000


Your leukapheresis sample is collected using ACD as the anticoagulant and
will contain ~300mg% citrate ion.  If you are experiencing coagulation, more
than likely your buffers/reagents contain divalent ions, specifically Ca++
and Mg++, thus causing the coagulation due to the binding to the citrate
ion.  Change to reagents without Ca++ and Mg++ and your problem should
disappear.

April

April G. Durett, MSc
Clinical Applications - Flow Cytometry
Center for Cell & Gene Therapy
Houston, TX




-----Original Message-----
From: Guo Yalin [mailto:GUO at mm11.ukl.uni-freiburg.de]
Sent: Thursday, September 28, 2000 6:04 AM
To: cyto-inbox
Subject: coagulation in leukapheresis



Dear flows,

My project is sorting SP cells, a kind of stem cells from human cord
blood, bone marrow and leukapheresis with Hoechst 33342. During fresh
leukapheresis cells were stained in Hoechst at 37  C, coagulation of
cells always happened. It's not good for both of staining and
sorting. I have tried with 1 mM EDTA and/or 1 U/ml DNase and didn't
get any good result. Does anybody has such experience and has some
solution to prevent the coagulation, which is definitely what I need?
I should appreciate if somebody could tell me in detail soon!

Thank you very much for any suggestion!

Yours sincerely,

Yalin Guo
Yalin Guo
Nothnagel Lab
Hematology/Onkology Department
University of Freiburg
Hugstetter Strasse 55
D-79106 Freiburg
Germany
Tel: +49 761 270 7199
Fax +49 761 270 7177
Email: guo at mm11.ukl.uni-freiburg.de



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