Isolation and cultivation of bone marrow-derived macrophages

Commonly used acronym: BMDM

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Scope of the method

The Method relates to
  • Animal health
The Method is situated in
  • Basic Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Animal derived cells / tissues / organs
Species from which cells/tissues/organs are derived
Mouse
Type of cells/tissues/organs
Bone marrow-derived macrophages

Description

Method keywords
  • macrophages
  • bone marrow
  • isolation
  • macrophage polarization
Scientific area keywords
  • Immunology
  • immunomodulation
  • Immunometabolism
  • macrophage polarization
Method description

Mononuclear cells are flushed from the bone marrow using a 25G needle. After centrifugation, the cells are resuspended in 50mL pre-heated DMEM containing 20ng/ml M-CSF. The cells are cultured in 10cm petri dishes for 7 days. At days 2,4 and 6, the medium is renewed after washing away non-adherent cells. On day 7, add 3mL enzyme-free dissociation buffer and suspend cells using a cell scraper. Transfer cells to a 15mL tube, centrifuge, count and seed for experiments.

Lab equipment
  • Flow cabinet ; 
  • Incubator ;
  • Centrifuge.
Method status
  • Published in peer reviewed journal

Pros, cons & Future potential

Advantages
  • Easy to use ; 
  • Reproducible.
Future & Other applications

Depending on the growth factors in the medium, different cell types can be cultured.

References, associated documents and other information

References

Lefere S, Van de Velde F, Hoorens A, Raevens S, Van Campenhout S, Vandierendonck A, Neyt S, Vandeghinste B, Vanhove C, Debbaut C, Verhelst X, van Dorpe J, Van Steenkiste C, Casteleyn C, Lapauw B, Van Vlierberghe H, Geerts A, Devisscher L. Angiopoietin-2 promotes pathological angiogenesis and is a novel therapeutic target in murine non-alcoholic fatty liver disease. Hepatology 2019 Mar;69(3):1087-1104.

Raevens S, Geerts A, Paridaens A, Lefere S, Verhelst X, Hoorens A, Van Dorpe J, Maes T, Bracke KR, Casteleyn C, Jonckx B, Horvatits T, Fuhrmann V, Van Vlierberghe H, Van Steenkiste C, Devisscher L, Colle I. Placental growth factor inhibition targets pulmonary angiogenesis and represents a novel therapy for hepatopulmonary syndrome in mice. Hepatology 2018 Aug;68(2):634-651.

Organisations

Ghent University (UGent)
Internal Medicine and Pediatrics
Belgium
Flemish Region

Partners

Ghent University hospital (UZ Gent)