Activation of Aged Murine BMDCs

The effects of combinations of polymeric nanoparticles, micelles, and various small molecule agonists on the activation of bone marrow derived DCs (BMDCs) harvested from the femur and tibia of aged (≥20 months) C57BL/6 male mice (n = 6) were studied. Bone marrow cells were cultured as outlined in Lutz et al. [101 (link)]. Briefly, cells were cultured in 10 mL RPMI 1640 medium supplemented with 100 U/mL penicillin, 100 mg/mL streptomycin, 2 mM glutamine, 10% FBS, and 20 ng/mL granulocyte-macrophage colony-stimulating factor (GM-CSF, Cat. #FB0875711Z, Peprotech, Rocky Hill, NJ) at approximately 5 × 10⁶ cells per 100 mm plate. On day 3 of culture, 10 mL of medium containing GM-CSF was added. On days 6 and 8 of the culture period, approximately half of the total volume of medium was removed and replaced with freshly supplemented RPMI. On day 10 plates were gently rinsed to harvest non-adherent DCs for assessment of activation and costimulatory expression. Animals were obtained from Jackson Laboratory (Bar Harbor, ME) and maintained at Iowa State University following IACUC protocol #IACUC-20-199.