Neutrophils were isolated from bone marrow of 4-week-old C57BL/6J mice. The femur and tibia were resected intactly and cut the two ends of the bone in a clean bench to flush the bone marrow out. After treatment with red cell lysis buffer, the neutrophils were isolated using Mouse neutrophil isolation kit (Miltenyi Biotec, 130–097-658) according to the manual. The isolated neutrophils were cultured in RPMI1640 medium supplemented with 10%FBS for future use.
Isolation and Characterization of Mouse Bone Marrow-Derived Mesenchymal Stem Cells and Neutrophils
Neutrophils were isolated from bone marrow of 4-week-old C57BL/6J mice. The femur and tibia were resected intactly and cut the two ends of the bone in a clean bench to flush the bone marrow out. After treatment with red cell lysis buffer, the neutrophils were isolated using Mouse neutrophil isolation kit (Miltenyi Biotec, 130–097-658) according to the manual. The isolated neutrophils were cultured in RPMI1640 medium supplemented with 10%FBS for future use.
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Corresponding Organization : Nanjing Drum Tower Hospital
Variable analysis
- Isolation of mouse bone marrow-derived MSCs from tibia and femoral marrow compartments
- Isolation of neutrophils from bone marrow of 4-week-old C57BL/6J mice
- Identification of mouse bone marrow-derived MSCs by flow cytometry
- Culturing of mouse bone marrow-derived MSCs in MSC culture medium
- Culturing of isolated neutrophils in RPMI1640 medium supplemented with 10% FBS
- Maintenance of cell incubator at 37°C with 5% CO2 and 95% humidity
- Changing of culture medium every 2-3 days
- Use of Trypsin-EDTA (0.25%) for passaging
- None specified
- None specified
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