Bone Marrow Harvest from Mouse Hind Limb

- Bone marrow is the spongy gelatinous tissue present in the hollow center of bones. It contains hematopoietic stem cells, which produce the blood cells, a key component of the immune system. Bone marrow isolation from larger bones, such as the femur and tibia, provides high cell yield.

To harvest bone marrow, place a euthanized mouse in a supine position and sterilize the skin using ethanol. Cut the abdominal cavity and remove the skin to expose the hind limbs. Break the tibia, a long bone of the leg, and place it in cold PBS to maintain the physiological conditions. Dislocate the kneecap and remove the muscles to expose the femur, the thigh bone.

Cut the femur off from the bottom of the bone and place in cold PBS. Punch a hole in a smaller volume microcentrifuge tube and place both bones in it with the bones' open-end facing the hole. Place the tube containing the bones into a larger volume microcentrifuge tube and centrifuge the double-layered tubes to pellet the bone marrow. Use the harvested bone marrow for further analysis. In this protocol, we will harvest bone marrow from the hind limbs of a mouse.

- For a bone marrow harvest, place a 6 to 10 week old C57 black-6 mouse in t