Bone marrow cells were isolated by flushing the femur and tibia of 8-week-old male C57BL/6 mice. The isolated bone marrow cells were incubated with macrophage colony-stimulating factor (M-CSF; Peprotech, Cranbury, NJ, USA), and were differentiated (7 days) into BMDMs. Bone marrow cells were incubated in RPMI medium containing 30 ng/ml M-CSF, 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 25 mM HEPES. The media were changed every 3 days.
Protocol detail
Mouse Bone Marrow-Derived Macrophage Isolation
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Bone marrow cells
C57bl mice
Femur
Fetal bovine serum
Hepes
M csf
Male
Penicillin
Streptomycin
Tibia
Corresponding Organization :
Other organizations : Seoul National University, Hanyang University, Daegu-Gyeongbuk Medical Innovation Foundation, Chosun University
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Variable analysis
independent variables
- Macrophage colony-stimulating factor (M-CSF) concentration
- Bone marrow cell differentiation into bone marrow-derived macrophages (BMDMs)
- Age of mice (8 weeks)
- Mouse strain (C57BL/6)
- Sex of mice (male)
- Culture medium (RPMI medium)
- Fetal bovine serum (FBS) concentration (10%)
- Penicillin/streptomycin concentration (1%)
- HEPES concentration (25 mM)
- Culture duration (7 days)
- Media change frequency (every 3 days)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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