A simple and effective method for the isolation and culture of human monocytes from small volumes of peripheral blood

Lara T Meital; Alexander S Coward; Mark Windsor; Tom G Bailey; Anna V Kuballa; Fraser D Russell

doi:10.1016/j.jim.2019.04.005

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A simple and effective method for the isolation and culture of human monocytes from small volumes of peripheral blood

Journal article

Peer reviewed

A simple and effective method for the isolation and culture of human monocytes from small volumes of peripheral blood

Lara T Meital, Alexander S Coward, Mark Windsor, Tom G Bailey, Anna V Kuballa and Fraser D Russell

Journal of Immunological Methods, Vol.472, pp.75-78

2019

DOI: https://doi.org/10.1016/j.jim.2019.04.005

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Abstract

monocytes

macrophages

cell isolation

Percoll®

Ficoll®

phagocytosis

Innate immune cell defects contribute to severe autoimmunity and the pathogenesis of inflammatory disease. Monocyte-derived macrophages typically retain disease related signatures and represent an excellent in vitro model to uncover and validate mechanisms contributing to specific pathological states. Monocyte isolation procedures vary widely in terms of purity, yield, cost, degree of technical difficulty and volume of peripheral blood needed. This paper outlines a novel isolation method that yields monocytes through density gradient centrifugation (Ficoll® and hyperosmotic Percoll®). The protocol has been optimised for small volumes of blood (42 ml) and is simple, reproducible and inexpensive compared to other methods. Monocyte recovery is 70% (relative to monocyte numbers within the buffy coat) and the highly functional macrophages produced are characterised by excellent purity (98.6 ± 0.6%) and intact activation and phagocytic capacities. The method is well suited to investigations involving patient populations where a particular subset of immune cells is known to contribute to the pathogenesis of a specific disease or is aberrant as a consequence of that disease.

Details

Title: A simple and effective method for the isolation and culture of human monocytes from small volumes of peripheral blood
Authors: Lara T Meital (Author) - University of the Sunshine Coast - School of Health & Sports Sciences
Alexander S Coward (Author) - University of the Sunshine Coast - School of Health & Sports Sciences
Mark Windsor (Author) - University of the Sunshine Coast - School of Health & Sports Sciences
Tom G Bailey (Author) - University of the Sunshine Coast - School of Health & Sports Sciences
Anna V Kuballa (Author) - University of the Sunshine Coast - School of Health & Sports Sciences
Fraser D Russell (Author) - University of the Sunshine Coast - School of Health & Sports Sciences
Publication details: Journal of Immunological Methods, Vol.472, pp.75-78
Publisher: Elsevier BV
Date published: 2019
DOI: 10.1016/j.jim.2019.04.005
ISSN: 0022-1759
Organisation Unit: School of Health - Biomedicine; UniSC Clinical Trials Centre; University of the Sunshine Coast, Queensland; School of Health and Sport Sciences - Legacy; School of Health - Sports & Exercise Science; Centre for Bioinnovation; School of Health and Behavioural Sciences - Legacy
Language: English
Record Identifier: 99451238802621
Output Type: Journal article

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Collaboration types: Domestic collaboration
Web Of Science research areas: Biochemical Research Methods; Immunology

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