Figure 1.

Isolation and extraction of bone marrow-derived macrophages

(A) Separate the entire thigh. i. Mice are sacrificed by CO₂ inhalation and soaked in 75% Ethyl Alcohol for 5 min. ii. Make an incision at the base of the thigh and expose the hind limbs by clipping outward with ophthalmic scissors. iii. Separate the entire thigh and remove the skin.

(B) Collect femurs and tibias. i. The separated thigh is washed with PBS containing P/S and placed in the biosafety cabinet. ii. Remove all muscle from the hind limbs and separate the joints to obtain femurs and tibias (achilles tendon, hip joint, and knee joint). iii. The ends of the femur and tibia are cut off to expose the bone marrow cavities. Scale bar: 1 cm or 4 cm.

(C) Flush out the bone marrow. i. The 1 mL of syringe needle (26G) is inserted into the bone marrow cavity, and the bone marrow cells are gently flushed from top to bottom into a 50 mL centrifuge tube. ii. The flushed cells are filtered through a 70 μm cell strainer.