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

• Published in: Cytokine (Philadelphia, Pa.) Vol. 61; no. 1; pp. 33 - 37
• Main Authors: Clanchy, Felix I.L., Hamilton, John A.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.01.2013
• Subjects: Antigens, CD34 - metabolism; CD34 antigen; Cell culture; Cell Differentiation; Cell Lineage; Cell Proliferation; Cells, Cultured; Culture Media, Serum-Free; cultured cells; Differentiation; FLT3L protein; Growth factors; Hematopoietic Stem Cells - physiology; Humans; Interleukin 3; Interleukin 6; Interleukin-3 - pharmacology; Interleukin-6 - pharmacology; Macrophage colony-stimulating factor; Macrophage Colony-Stimulating Factor - pharmacology; Macrophages; Macrophages - metabolism; Membrane Proteins - pharmacology; Monocytes; Monocytes - metabolism; Peripheral blood; Phagocytosis; phenotype; Serum-free medium; Stem Cell Factor - pharmacology; Stem cells; Surface markers; Serum-free medium; MDM; Stem cells; PDM; Mφ; Macrophages; Growth factors; DC
• ISSN: 1043-4666; 1096-0023
• DOI: 10.1016/j.cyto.2012.09.005

► Serum-free cultures of human CD34+ progenitors are differentiated to macrophages. ► Optimal expansion of cell numbers was obtained by the presence of IL-3, SCF, IL-6 and M-CSF. ► The phenotype of progenitor-derived macrophages is similar to monocyte-derived macrophages. 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.