Murine Palatal Shelf Explant Culture

Pregnant mice were euthanized on day 14 of gestation (E14.0) under ketamine (25 mg/kg)/Rompun (8 mg/kg) anesthesia using sterile conditions, and the fetuses were removed from the uterus and placed in BGJb medium (gibco@ Life technologies). Palatal explants were dissected under a dissection microscope as described previously (Charoenchaikorn et al., 2009 (link)). Dissected palatal shelves were cultured in a glass bottom dish (Matsunami, Osaka, Japan) with medium containing 0.6% low melting agarose (Wako Osaka, Japan) (Kim et al., 2015 (link)). The medium was 500 μl of BGJb supplemented with 100 μg/ml penicillin/streptomycin (Invitrogen). Explants were cultured at 37°C with 5% CO₂ using a standard CO₂ incubator for 20 h.