In our hands, the Hoechst method is relatively specific for the early stem
cell. There are literature reports of cells identified by the Rhodamine
123 technique that have progenitor cell properties. Part of our current
investigations relate to the actual mechanism of the specificity. Data
from the ASH meeting indicate the cells do indeed take up the Hoechst dye,
but the "Side population cells" appear to exclude the dye from the nucleus.
This could indicate a pump or port on the nuclear membrane is responsible
for the failure of the Hoechst to stain the DNA in the nucleus.
Bill Telford
<telfordw@box- To: Cytometry Mailing List
<cytometry@flowcyt.cyto.purdue.edu>
t.nih.gov> cc:
Subject: Side population detection
01/03/2001
09:15 AM
Hello everyone...
We have a group at NIH doing side population analysis using Hoechst 33342
by the standard technique. My (probably obvious) question is...is there
something special about the Hoechst dye (mdr type specificity, nuclear
localization, etc) that makes it the only mdr susbtrate effective for stem
cell identification? Is it possible to use other fluorescent mdr
substrates (i.e. rhodamine 123, daunomycin, etc.) to identify these cells?
Thanks in advance!
Bill Telford
DCS-NCI-NIH
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