Murine Dendritic Cell Generation

Six-week-old female C57BL/6 mice were purchased from Orient Bio (Gyeonggi-do, Korea). Six-week-old female wild-type C3H/HeN and congenic TLR4-defective C3H/HeJ mice were purchased from Central Lab. Animal Inc. (Seoul, Korea). All animals were housed in a specific pathogen-free (SPF) facility at the Korea Research Institute of Bioscience and Biotechnology (KRIBB). All experiments employing mice were reviewed and approved by the Institutional Animal Care and Use Committee of the KRIBB. Immature BMDCs were generated using Roswell Park Memorial Institute 1640 media (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Gibco), 100 U/mL penicillin, 100 mg/mL streptomycin (Gibco), 20 ng/mL murine granulocyte-macrophage colony-stimulating factor (GM-CSF; Peprotech), and 10 ng/mL murine IL-4 (Peprotech) for 7 days followed by changing with fresh media every 3 days, as previously described [21 (link),22 (link)].