Expression of the macrophage-specific antigen F4/80 during differentiation of mouse bone marrow cells in culture.

Abstract

Cells of the mononuclear phagocyte system develop from precursors in the bone marrow (1), but little is known about their pathway of differentiation or its control in vivo. The development of morphology (2) (8), which are found on mouse m~ but not on other hemopoietic or unrelated cells, provide new markers to define stages in the differentiation of m~b and to characterize and isolate their precursors.In this paper, we analyze the expression of Ag F4/80 on cells of the m~ lineage from progenitors in bone marrow to mature ceils obtained after culture in the presence of L cell-conditioned medium (LCM). F4/80 is also used in a clonal approach to examine mq~ subset heterogeneity. Materials and MethodsAnimals. Swiss mice and CBA T6T6 mice of both sexes were bred in this department. Similar results were obtained using either sex or strain.Cells. Bone marrow (BM) and resident peritoneal cells were obtained as described (7,8). Mass liquid cultures of BM were prepared in a BM culture medium (BMCM) consisting of Dulbecco's modification of minimal essential medium (DMEM) (high glucose), 15% heatinactivated (HI) horse serum (Flow Laboratories, Inc., Irvine, Scotland), 1% HI fetal bovine serum, 2 mM glutamine, 100 U/ml penicillin, 100 #g/ml streptomycin, I00/xg/ml kanamycin, and 10-20% LCM. BM 2 cells were cultured in 75-or 175-cm tissue culture flasks (Falcon Labware, Div. of Becton, Dickinson & Co., Oxnard, Calif., and Nunclon, Gibco, Middlesex, England.) 25 or 50 ml, respectively, at 8 × 10 n nucleated cells/ml. Flasks were coated with gelatin as described (7).Cultures were divided into a nonadherent cell (NAC) fraction, usually after 3 d, and adherent

Morphological characterization of stromal cell types in hematopoietically active long-term murine bone marrow cultures.

“…The macrophage progenitors (CFU-M in cultures) lack the F4/80 antigen, whereas macrophages in formed colonies are positive. The adherent macrophages expressed high level of F4/80 antigen (35). The MAb F4/80-positive cells were polygonal, stellate, and occasionally bipolar.…”

Differential Ly6C Expression after Renal Ischemia-Reperfusion Identifies Unique Macrophage Populations

“…PCNA staining and vimentin expression peak at this time ( Figure 1D), suggesting this population may be involved in repair. Finally, the Ly6C 27 Moreover, inhibition of the c-fms receptor which mediates proliferation, differentiation and survival of monocytes/ macrophages 28 when bound to colony-stimulating factor demonstrates that inhibition of monocytes/macrophages prior to IRI ( Figure 3A) dampens the proinflammatory milieu leading to renoprotection. An approximately 50% decrease is measured 2 hours postreperfusion in kidney mRNA expression of IL-6 (Figure 3F) and CXCL1 ( Figure 3G), with significant reductions in IL-6 at 1 day.…”

The Induction of Inflammation by Dectin-1 In Vivo Is Dependent on Myeloid Cell Programming and the Progression of Phagocytosis

“…Bone marrow-derived MØ (BMMØ) and BMDC were produced by culture of bone marrow cells from the femur and tibia of mice in the presence of M-CSF or GM-CSF (both Peprotec), respectively, for 7-10 days as previously described (31,32).…”