Bone Marrow-Derived Dendritic Cells Generation: A Method to Generate Dendritic Cells from Mouse Bone Marrow

- Dendritic cells or DCs are key antigen presenting cells that activate T-cells. In leukemic patients, these cells poorly express tumor associated antigen and fail to elicit an antigen specific immune response. Dendritic cells are abundant in the bone marrow. And hence, the bone marrow is the preferred site for their isolation to help study their role in cancer.

To obtain dendritic cells, extract the bone marrow from a euthanized mouse's femur or tibia bone by using a media filled syringe to flush it out. Centrifuge and resuspended the pellet in ammonium-chloride-potassium or ACK buffer to lyse erythrocytes. Now add culture media to stop lysis and centrifuge. Resuspend the pellet in the culture media containing an antibiotic. Add granulocyte-macrophage colony-stimulating factor or GM-CSF, and transfer the cell suspension to a Petri plate.

Incubate the plate for the desired time duration. GM-CSF helps progenitor cells to differentiate into dendritic cells. Finally, activate the cells with lipopolysaccharides or LPS, which binds to the surface receptor of dendritic cells, resulting in fully mature and immunogenic bone marrow dendritic cells. In this protocol, we will demonstrate the