Published online before print January 24, 2006

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(Journal of Leukocyte Biology. 2006;79:757-766.)
© 2006 by Society for Leukocyte Biology

Detection and properties of the human proliferative monocyte subpopulation

Felix I. L. Clanchy^*,, Alice C. Holloway^*, Roya Lari^*,, Paul U. Cameron^,1 and John A. Hamilton^*,,2

^* Departments of Medicine, Royal Melbourne Hospital, and
Microbiology and Immunology, University of Melbourne, Parkville, Victoria, Australia; and
Cooperative Research Centre for Chronic Inflammatory Diseases, Parkville, Victoria, Australia

²Correspondence: University of Melbourne, Department of Medicine, Royal Melbourne Hospital, Parkville, Victoria, 3050, Australia. E-mail: jahami{at}unimelb.edu.au

Peripheral blood monocyte subpopulations have been reported and can give rise to diverse, differentiated phenotypes. A subpopulation(s) of human monocytes can proliferate in vitro in response to macrophage-colony stimulating factor (M-CSF; or CSF-1). This population, termed the proliferative monocyte (PM), is presumably less mature than other monocytes; however, it has not been defined further. Previous studies monitoring the frequency of the slowly cycling PM from different donors indicated that the assay for their reproducible measurement required improvement. We demonstrate that for optimal PM detection, high 5-bromo-2'-deoxyuridine concentrations are required over a delayed and wide time-frame. Surface marker phenotyping by flow cytometry showed that freshly isolated PM are CD14⁺ and could be distinguished from two other human monocyte subpopulations, namely, the CD14^loCD16⁺ and CD14^loCD64^– subsets. PM express relatively high levels of CD64 and CD33 but have relatively low CD13 expression; they are also c-Fms⁺ and human leukocyte antigen-DR⁺. Labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) enabled the estimation of the number of PM divisions over time. Following CFSE labeling and culture, PM were sorted from the nonproliferating population and shown to have a distinctive, spindle-shaped morphology and higher capacity to form multinucleated, tartrate-resistant acid phosphatase⁺ cells in the presence of M-CSF and receptor activator of nuclear factor-B ligand. The phenotype and properties of the PM subpopulation were examined as a prelude to determining its role in disease using methods that can be applied to clarify human monocyte heterogeneity.

Key Words: macrophages • cell proliferation • cell differentiation

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