degranulation

From: Arnold Pizzey (a.pizzey@ucl.ac.uk)
Date: Fri Mar 16 2001 - 07:28:17 EST


>Hi, I have a question.  The question is: What do people do to prevent
>peripheral blood neutrophils from degranulating during handling and
>storage. Are there special reagents, media, or buffers that can be used?
>Thank you in advance,
>
>Victoria Boryca Smith
>UNMC
>vbsmith@unmc.edu


Hello Victoria,

Try Dulbeco's PBS +Ca/Mg 5mM Glucose *NO FCS* cells at~ 1exp6/ml and keep
the cells at Room Temp (NOT on ice)
I've used the above conditions to maintain PMNs for ~6 hours and the
viability is good.
What is more important is, I believe the method by which you obtained the
cells in the first place. Prolonged contact with Dextran/Ficoll tends to
get the cells reved up -I have my own method for isolation which minimises
contact times -if you contact me directly I'll pass it on.

Of course if you are not bothered about cell viability you can just throw
truckloads of DIFP/DMEM at them

Regards,

Arnold


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	Arnold Richard Pizzey
	Department of Haematology
	Royal Free and University College London Medical School
	98 Chenies Mews
	London WC1E 6HX
	U.K

	voice:	+44 (0)20-7679-6234
	Fax:	+44 (0)20-7679-6222
	email:	a.pizzey@ucl.ac.uk
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