Characterization of subsets of bone marrow-derived macrophages by flow cytometry analysis

Journal Article · Tue Jan 01 04:00:00 EST 1985 · J. Leukocyte Biol.; (United States)

OSTI ID:5819563

Walker, E B; Akporiaye, E T; Warner, N L; Stewart, C C

Normal C3H bone marrow cells were grown 7 days in medium containing L cell-derived colony stimulating factor-1 (CSF-1). During the first 4 days of culture, erythroid and granulocytic cells decreased while macrophages increased exponentially with a doubling time of about 31 hr. Only 0.3% of all cells in the initial bone marrrow suspension formed discrete colonies of mononuclear phagocytes, but by day 6 60% of the nonadherent cells were capable of forming macrophage colonies, representing a 200-fold enrichment of the original progenitor population. Using flow cytometry, mononuclear phagocytes obtained after 4 days of culture were separated into two distinct phenotypes based on their autofluorescence. Nonadherent cells were a discrete population of small cells exhibiting low autofluorescence, and the adherent cells were a broad heterogenous population of large cells exhibiting high autofluorescence. A panel of currently available rat monoclonal antibodies (MABs) against murine hematopoietic cells were used to determine whether unique subsets of macrophages could be resolved. The MABs RA 31B6 and H-11 stained virtually all the nonadherent cells but not adherent cells. The MABs E-2 and 11-4.1 (anti-H-2K/sup k/) stained almost all the adherent cells and demonstrated no significant staining of nonadherent cells. Nearly all the nonadherent and adherent cells were stained by the MABs DNL 4.4 and MAC-1. Additionally, the data suggest that the epitopes for MAC-2 and MAC-3 and ..gamma..2a Fc receptors develop late in nonadherent progenitor cells as they mature into adherent macrophages. 37 references, 5 figures, 5 tables.

• Research Organization: Los Alamos National Lab., NM
• OSTI ID: 5819563
• Journal Information: J. Leukocyte Biol.; (United States), Journal Name: J. Leukocyte Biol.; (United States) Vol. 37; ISSN JLBIE
• Country of Publication: United States
• Language: English

Related Subjects

550300* -- Cytology
59 BASIC BIOLOGICAL SCIENCES
ADHESION
ANIMAL CELLS
ANIMALS
ANTIBODIES
BONE MARROW CELLS
CELL CULTURES
CELL FLOW SYSTEMS
COLONY FORMATION
CONNECTIVE TISSUE CELLS
DATA
EXPERIMENTAL DATA
FLUORESCENCE
GROWTH
INFORMATION
LUMINESCENCE
MACROPHAGES
MAMMALS
MONOCLONAL ANTIBODIES
NUMERICAL DATA
PHAGOCYTES
RATS
RODENTS
SOMATIC CELLS
STAINS
VERTEBRATES