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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.

Original publication

DOI

10.1016/j.cyto.2012.09.005

Type

Journal article

Journal

Cytokine

Publication Date

01/2013

Volume

61

Pages

33 - 37

Addresses

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

Keywords

Monocytes, Hematopoietic Stem Cells, Cells, Cultured, Macrophages, Humans, Interleukin-3, Macrophage Colony-Stimulating Factor, Membrane Proteins, Antigens, CD34, Stem Cell Factor, Interleukin-6, Culture Media, Serum-Free, Cell Differentiation, Cell Proliferation, Cell Lineage