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We have looked at HUVEC and other adherent cell lines by flow quite a lot.
The other population you mentioned is very likely aggregates. I would
recommend using Sigma's Cell Dissociation solution (cat #C-5914) instead of
trypsin to remove the cells from the plates or flasks. Pipette them up and
down 8-10 times to get a good single cell suspension and then wash. After
washing, pour them through a Falcon 100uM cell strainer (cat. # 2360). This
usually gives a fairly homogenous-looking population by scatter and you won't
lose cell surface antigens to the trypsin.
Gary Elliott
AMGEN Flow Cytometry
--------------------------------------
Date: 7/3/96 12:42 PM
To: cyto-inbox
From: DODONNEL@svherc.ucd.ie
Has anybody out there any experience looking at endothelial cells ? A
researcher in our lab asked me to look at some of hers (HUVEC line) with a
view to examining expression of adhesion markers in the future.
Although on microscopy they seemed to be a population of fairly large cells
of quite uniform size, on the FSC/SSC plot there seemed to be 2 populations,
one with moderate to high FSC and SSC increasing proportionately,
the other with a lower SSC but an FSC going off the scale.
I wondered if the 2nd lot represented aggregated cells, although she did
trypsinise the cells before fixing them. Both populations had the same
autofluorescence characteristics.
I'd be very grateful for any words of wisdom.
Dr. Dearbhaile O' Donnell
University College Dublin
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Date: Wed, 03 Jul 1996 15:06:39 +0100 (BST)
Subject: Endothelial cells by flow
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