Identification and isolation of splenic tissue-resident macrophage sub-populations by flow cytometry

• Published in: International immunology Vol. 31; no. 1; pp. 51 - 56
• Main Authors: Fujiyama, Satoshi, Nakahashi-Oda, Chigusa, Abe, Fumie, Wang, Yaqiu, Sato, Kazuki, Shibuya, Akira
• Format: Journal Article
• Language: English
• Published: UK Oxford University Press 06.02.2019
• Subjects: Animals; Biomarkers; Cell Separation - methods; Flow Cytometry - methods; Immunohistochemistry; Immunophenotyping; Macrophages - immunology; Macrophages - metabolism; Mice; Phagocytosis; Short Communication; Spleen - cytology; Spleen - immunology; Spleen - metabolism; enzymatic dissociation; marginal zone macrophages; marginal zone metallophilic macrophages
• ISSN: 0953-8178; 1460-2377
• DOI: 10.1093/intimm/dxy064

Isolation of splenic macrophage subpopulations using an enzyme cocktail Abstract Tissue-resident macrophages in the spleen, including red pulp and white pulp macrophages, marginal zone macrophages (MZMs) and marginal zone metallophilic macrophages (MMMs), are highly heterogeneous as a consequence of adaptation to tissue-specific environments. Each macrophage sub-population in the spleen is usually identified based on the localization, morphology and membrane antigen expression by immunohistochemistry. However, their phenotypical and functional characteristics remain incompletely understood due to the difficulty of identification and isolation by flow cytometry. We used a cocktail of three enzymes (Collagenase D, Dispase I and DNase I), rather than traditional mechanical grinding, for isolation of each sub-population, which resulted in significant improvement of isolation of these macrophage sub-populations, particularly MZMs and MMMs, as determined by CD11bhiF4/80medTim4hi and CD11bhiF4/80medTim4med, respectively. This method should be helpful for molecular and functional characterization of each splenic resident macrophage sub-population.