Bone Marrow Derived Macrophage (BMDM) differentiation and maintenance
Narayana Yadavalli, Shawn M. Ferguson
Published: 2023-08-05 DOI: 10.17504/protocols.io.j8nlkw7bwl5r/v1
Abstract
This protocol describes the isolation of bone marrow derived macrophages.
Attachments
Steps
1. Note : For mouse bone marrow-derived macrophage (BMDMs) primary cultures, each experiment involved age and sex matched C57BL/6 mice between 3-9 months of age. Lrrk2 KO (Lrrk2tm1.1Mjff) and G2019S (Lrrk2tm1.1Hlme) homozygous knockin mice were obtained from The Jackson Laboratory76. Euthanize mice via CO2 or isoflurane inhalation and cervical dislocation.
Note
2.
Collect femurs and flush cavities with 5mL ice-cold PBS.
3.
Then, collect bone marrow cells by centrifugation 0.3rcf.
4.
Resuspend the resulting pellet and differentiate for 6 days in culture media containing: DMEMF12 supplemented with 20% FBS, 20% L929 conditioned media, 1% penicillin-streptomycin and 1% GlutaMAXTM.
