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Cytokine. 2013 Jan;61(1):33-7. doi: 10.1016/j.cyto.2012.09.005. Epub 2012 Oct 2.

The development of macrophages from human CD34+ haematopoietic stem cells in serum-free cultures is optimized by IL-3 and SCF.

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  • 1Arthritis and Inflammation Research Centre, Department of Medicine, Royal Melbourne Hospital, University of Melbourne, Parkville, Victoria, Australia. felix.clanchy@kennedy.ox.ac.uk

Abstract

The derivation of human macrophages from peripheral blood monocytes remains a convenient method for the study of macrophage biology. However, for macrophage differentiation, a significant proportion of development has occurred prior to the monocyte stage; monocyte subsets also have varying potential for differentiation. Differentiation of macrophages from a less mature precursor, such as CD34+ haematopoietic stem cells, can further inform with regard to the development of macrophage-lineage cells. CD34+ cells were cultured in serum-free medium containing Flt3L, SCF, IL-3, IL-6 and M-CSF. Using differing combinations of growth factors, the effect on cell proliferation and differentiation to adherent macrophage-like cells was determined. The proliferative response of CD34+ cells to M-CSF was determined during the initial phase of cell culture. Thirteen combinations of SCF, IL-3, IL-6 and M-CSF were then compared to determine the optimum combination for proliferation. Adherence was used to isolate mature macrophages, and the macrophage-like phenotype was confirmed by analyses of surface markers, histo-morphology and phagocytosis. This study refines the means by which large numbers of macrophages are obtained under serum-free conditions from haematopoietic precursors.

Copyright © 2012 Elsevier Ltd. All rights reserved.

PMID:
23040055
[PubMed - indexed for MEDLINE]
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