Re: G0/G1 and FITC

STEVEN DAVID GORE (sdgore@welchlink.welch.jhu.edu)
Thu, 25 May 1995 08:11:07 -0400 (EDT)

On Wed, 24 May 1995 bpijb@picr.cr.man.ac.uk wrote:

> One of our researchers has expressed the desire to analyse
> and then sort G0 phase cells stained with FITC (possibly PE) derived
> from Cord/Bone Marrow samples.

Two possibilities: if you don't need the cells to be viable, Ki67
staining works well on bone marrow and cord blood, and can be used in
double label experiments with CD34 and other surface markers. Note,
however, that the lack of expression of Ki67 in G0 cells was evaluated by
Gerdes in resting lymphocytes versus PHA-stimulated lymphocytes. We have
used Ki67 to enumerate "G0" cells and follow proliferation in normal CD34
cells as well as leukemic cells, eg:

Exp. Heme. 23:413-421, 1995
Exp. Heme 21: 1702-1708, 1993.

The latter paper is more methodological.

The other choice, if you want viable cells, would be Rhodamine 123. I do
not have personal experience with Rhodamine, but there is an extensive
literature out there: one early reference Ploemacher and Brons, J. Cell
Phys 136:531-536, 88.

Steve Gore
Johns Hopkins Oncology Center


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