Bone Marrow-Derived Macrophages isolation and differentiation
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Animal derived cells / tissues / organs
- macrophage polarization
Isolation Femoral and tibial bones are isolated from 10-12 weeks old C57BL/6J mice. After cutting the bone's ends, the bone marrow is flushed out using a 23G syringe with PBS into a 50ml falcon tube. After centrifugation cells are re-suspended in FCS (0.5ml/mouse leg). Differentiation 0.5ml of the cells suspension is added to 20ml BMM medium (15% LCM, 10% FCS and 0.5% P/S in RPMI1640) in a 20cm petri dish. After 5 days of culture all medium is replaced by 20ml fresh BMM medium. On day 7 cells can be plate out for experiments.
- - Biosafety cabinet flow hood ;
- - 5% CO2 Incubator ;
- - Centrifuge ;
- - Dissection material.
- History of use
- Internally validated
- Published in peer reviewed journal
Pros, cons & Future potential
- - High amount of cells per isolation ;
- - Possibility of freezing the cells for future use after isolation ;
- - Reproducibility.
If the cells are confluent at day 5, instead of discard the BMM medium you can place it into a new petridish to increase the amount of cells.
References, associated documents and other information
Bogie JF, Mailleux J, Wouters E, Jorissen W, Grajchen E, Vanmol J, Wouters K, Hellings N, Van Horsen J, Vanmierlo T, et al. 2017b. Scavenger receptor collectin placenta 1 is a novel receptor involved in the uptake of myelin by phagocytes. Sci Rep 7: 44794.
Contact personJerome Hendriks