Badley, Andrew abadley at ohri.ca
Tue Mar 19 16:14:35 EST 2002

Positive selection by CD14 can result in non physiologis activation of the
monocyte ( presumably akin to LPS stiumulation) which in our hands does
result in monocyte death. I am unsure why negative selection would also
result in death.
One possibility might be the length of time samples are run during the
selection process. Have you tried bead selection to remove CD3, 19, 56,
followed by a flow separation to ensure purity??

Andrew Badley MD, FRCPC
Assistant Professor, Division of Infectious Diseases
Ottawa Hospital, General Campus
501 Smyth Road
Ottawa, Ontario CANADA

Fax 613-737-8682

> -----Original Message-----
> From: Kraus, Elizabeth [SMTP:elizabkr at bhcs.com]
> Sent: Monday, March 18, 2002 1:05 PM
> To:	Cytometry Mailing List
> Subject:	CELL DEATH
> For those of you with an AutoMACS in your facility, please allow me to
> pick your brains.
> We have been performing either negative selections or positive
> depletions to enrich for monocytes on a routine and successful basis.
> Suddenly, our cells are dying after 1-2 days in culture.  Using the same
> starting sample, cells isolated in parallel but by adherence are doing
> fine.  Naturally, all reagents have been tossed and re-made but still no
> change.  Additionally, there have been no changes to the AutoMACS
> cleansing reagents (just EtOH and bleach as recommended by Miltenyi) and
> the instrument is clean as a whistle.  Any suggestions would be
> appreciated.	I know this is vague but I'm crossing my fingers that
> someone out there can shed some light.
> Elizabeth Kraus
> B.I.I.R.
> Dallas, TX.

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