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J Vis Exp. 2014 Dec 27;(94). doi: 10.3791/52347.

Isolation and intravenous injection of murine bone marrow derived monocytes.

Author information

1
Department for Cardiology, Angiology and Pneumology, Otto von Guericke University Magdeburg.
2
Herzzentrum Dresden, Universitätsklinikum an der Technischen Universität Dresden, Technische Universität Dresden.
3
Department of Public Health and Primary Care, University of Cambridge.
4
Department for Cardiology, Angiology and Pneumology, Otto von Guericke University Magdeburg; joerg.herold@med.ovgu.de.

Abstract

As a subtype of leukocytes and progenitors of macrophages, monocytes are involved in many important processes of organisms and are often the subject of various fields in biomedical science. The method described below is a simple and effective way to isolate murine monocytes from heterogeneous bone marrow. Bone marrow from the femur and tibia of Balb/c mice is harvested by flushing with phosphate buffered saline (PBS). Cell suspension is supplemented with macrophage-colony stimulating factor (M-CSF) and cultured on ultra-low attachment surfaces to avoid adhesion-triggered differentiation of monocytes. The properties and differentiation of monocytes are characterized at various intervals. Fluorescence activated cell sorting (FACS), with markers like CD11b, CD115, and F4/80, is used for phenotyping. At the end of cultivation, the suspension consists of 45%± 12% monocytes. By removing adhesive macrophages, the purity can be raised up to 86%± 6%. After the isolation, monocytes can be utilized in various ways, and one of the most effective and common methods for in vivo delivery is intravenous tail vein injection. This technique of isolation and application is important for mouse model studies, especially in the fields of inflammation or immunology. Monocytes can also be used therapeutically in mouse disease models.

PMID:
25591000
PMCID:
PMC4354490
DOI:
10.3791/52347
[Indexed for MEDLINE]
Free PMC Article

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