Isolation and Differentiation of Murine Bone Marrow-Derived Macrophages
Bone marrow cells were collected from femurs and tibia of both male and female 6‐ to 8‐week old C57BL/6J or C57BL/6NTac wild‐type mice. Collected cells were treated with red blood cell lysis buffer (155 mM NH4Cl, 12 mM NaHCO3, 0.1 mM EDTA) and plated on untreated 10 cm cell culture dishes (BD Biosciences) in IMDM (Gibco) containing 10% heat‐inactivated FBS, 100 units/ml penicillin/streptomycin (Gibco) and 15% L929 conditioned supplement. After 24 h, the cells in supernatant were transferred to untreated 10 cm Petri dishes (BD Biosciences) for 7 days for the differentiation into bone marrow‐derived macrophages (BMDMs) (Heap et al, 2021 (link)).
Bilkei‐Gorzo O., Heunis T., Marín‐Rubio J.L., Cianfanelli F.R., Raymond B.B., Inns J., Fabrikova D., Peltier J., Oakley F., Schmid R., Härtlova A, & Trost M. (2022). The E3 ubiquitin ligase RNF115 regulates phagosome maturation and host response to bacterial infection. The EMBO Journal, 41(23), e108970.
Bone marrow cells Bone marrow derived macrophages Buffer C57bl mice Cell culture Dishes Edta Female Femurs Male Nahco3 Penicillin Red blood cellStreptomycin SupplementTibia
Corresponding Organization :
Other organizations :
MRC Protein Phosphorylation and Ubiquitylation Unit, University of Gothenburg, University of Dundee, Newcastle University, University of Leicester
Bone marrow-derived macrophage (BMDM) differentiation
control variables
Age of mice (6‐ to 8‐week old)
Cell culture media (IMDM containing 10% heat‐inactivated FBS, 100 units/ml penicillin/streptomycin, and 15% L929 conditioned supplement)
Cell culture dishes (untreated 10 cm cell culture dishes and Petri dishes)
controls
Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.
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