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We routinely quantitate CD34+ stem cells in peripheral blod and stem cell
harvest products.
We use a very simple method: whole blood or cell product is labelled with
BDs anti-CD34-PE antibody conjugate, lysed and then acquired on a FACScan
with threshold set on forward scatter. Stem cells are determined as CD34+
cells with low side scatter. Absolute WBC counts are determined on a
hematology counter (Sysmex NE 8000).
We are very satisfied with the method. The stem cell cluster is easily
determined, and the predictability on the stem cell count in the stem cell
product is quite good. We find, that using a second (eg anti-CD45)
antibody complicates things, without giving any extra information.
Normally, we see higher CD34+ percentages when using a CD34 CD45 dual
color protocol.
A workshop for Nordic stem cell transplantation centres held i 1995 in
Herlev, Denmark, in fact revealed, that the major problem concerning
laboratory to laboratory variation in determining absolute CD34+ counts, is
depending more on the method used for WBC counts, than the method used for
flow cytometric deternmination of the CD34+ percentage.
Let me know if you would want a more detailed version of our protocol.
Ulrik Sprogoe-Jakobsen
Dept. Clinical Immunoloy
Odense University Hospital
Denmark
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web