viability -Reply

From: Mike Keeney (Mike.Keeney@LHSC.ON.CA)
Date: Thu Jan 07 1999 - 09:03:06 EST


Hi Cheryl,

There are numerous potential reasons why your viability may be low on
peripheral apheresis collections. How is viability being measured? Is it
the actual CD34 cells that are non viable or the pack viabilty in general
which is low? Here are a few things which may be affecting your
viability:
Granulocyte contamination - granulocytes die within a few hours of
collection, especially if packs are stored at room temperature with high
cell concentrartions. 
Time from collection to analysis - Were the samples run fresh on these
patients or stored overnight and run the next day? I am always amazed
at how few people realise that most flow labs don't run 24 hours a day
and often store samples overnight. If samples are stored overnight at
room temperature the viabilty is reduced significantly, however if stored
at 4C the viability is well maintained (usually >95%).
Is viabilty measured at the time of analysis in a flow assay? If this is the
case and 7-AAD or PI is used the time from sample prep, the lysing agent
and storage conditions will all impact viabilty. Samples lysed in ammonium
chloride must be held ON ICE for a maximum of 1 hour before flow
analysis. 
I hope some of these ideas are helpful. 

Mike

Michael Keeney
Technical Specialist
Hematology/Flow Cytometry
London Health Sciences Centre
800 Commissioners Road East
London, Ontario
Canada
N5X 2Z2

>>> "Cheryl Hardy" <chardy@medicine.umsmed.edu> 01/05/99 05:13pm
>>>

Could someone please give me an explanation as to why CD34 cell
viability would be lower than normal on a particular patient's apheresis
collection? Our FLOW unit provides a viability based upon PI staining
for each CD34 it runs for our BMT transplant program, and they seldom
are below 95%, as they are always run immediately upon delivery to the
FLOW lab. Then, a patient came along with 70% viability one day on his
apheresis collection and 84% the following day. Thoughts?

Thanks in advance,
Cheryl Hardy


Cheryl L. Hardy, Ph.D.
Associate Professor of Medicine (Research)
Univ. of Mississippi School of Medicine
2500 N. State St.
Jackson MS 39216

601-984-6293
601-984-5689 FAX



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