Citrate vs. CTAD in platelet activation studies

From: Michael F. Wolf (michael.wolf@medtronic.com)
Date: Thu Oct 16 1997 - 05:04:59 EST


Some articles in the literature indicate that blood anticoagulant type
and amount play a role in expression of certain cell activation markers
e.g.,  expressed on WBCs and platelets (I'm interested in any references
here - along with info on the Q below).  In my particular in vitro
studies,  however, I often try to mimic clinical CPB (cardiopulmonary
bypass) conditions using human whole blood (50-100% diluted) and heparin
as anticoagulant.  The latter also includes dose response analyses
e.g.,  with subclinical (e.g., << 1 U/ml) heparin levels.  My question
for anyone out there is: as I need to immediately quench generation of
coagulation factors (particularly thrombin generation,  also platelet
activity etc..) at specified timepoints in the studies,  what's the best
anticoagulant or other agent for this?  Certainly I could just use
fixative.  The BD 3-color procedure suggests ACD but CTAD seems like it
would be a good choice too - because of the presence of platelet
stabilizers against further invitro platelet activation.  To complicate
things,  I also run quite a few ELISAs (TAT,  C3a,  etc..) on the
collected blood - so I have to be cautious about interfering agents.



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