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Characterization of Subsets of Bone Marrow-Derived Macrophages by Flow Cytometry Analysis

生物 骨髓 流式细胞术 人口 分子生物学 单克隆抗体 造血 祖细胞 巨噬细胞 外周血单个核细胞 免疫学 自体荧光 抗体 病理 干细胞 细胞生物学 体外 医学 生物化学 物理 环境卫生 荧光 量子力学
作者
Edwin Walker,Emmanuel T. Akporiaye,Noel L. Warner,Carleton C. Stewart
出处
期刊:Journal of Leukocyte Biology [Oxford University Press]
卷期号:37 (2): 121-136 被引量:53
标识
DOI:10.1002/jlb.37.2.121
摘要

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 marrow 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 heterogeneous 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-2Kk) 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.

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