On Tue, 25 Mar 1997 "RICHARD L. DARLEY" asked > > I have been carrying out culture of CD34+ cells in the presence of > IL-3, GM- & G-CSF and following myeloid differentiation using cell > surface markers. > > I am finding that staining with CD14 is grossly under-estimating the > monocytic content of these cultures as judged by morphology. > > One of my colleagues has made similar observations. Additionally he > found that CD14+ AML cells lost expression of this marker when in > culture. > > Has anyone had similar experiences or any good suggestions for > alternative markers which more reliably indicate monocytic > differentiation under these conditions? > > CD14 will stain a subset only even of peripheral blood monocytes. These cells are extremely heterogeneous and multi-color analysis will reveal at least five subpopulations (Arterioscler Thromb Vasc Cell Biol 16: 1437-47, 1996). To our knowledge there is no surface marker available which will stain all monocytes. CD64 is similar in expression to CD14 but is also expressed on activated neutrophils. CD33 will detect more monocytes but all neutrophils will also stain dimly. The ideal marker would be CD68 but I am not aware of a good commercial clone which will react with the extracellular portion of this antigen. Gregor Rothe ************************************************************************* Institute for Clinical Chemistry and Laboratory Medicine University of Regensburg D-93042 Regensburg, Germany Tel. +49 (941) 944-6204 Fax +49 (941) 944-6202 Internet: Gregor.Rothe@klinik.uni-regensburg.de *************************************************************************
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