Feather Buds Open blunt end of chick egg. Dump contents into large petri dish. Transfer embryo to clean 60 mm dish with sterile Howard's. Remove head close to shoulders and discard. Use fine forceps to slice along flank; try to only cut through the outer layer (skin). Using fine forceps gently peel back skin, beginning at the neck. Try to remove a large intact piece.   Skin media: Dulbecco's modified Eagles medium, 10% fetal calf serum, 50 ug/ml gentamicin   Lung Primordia Dissect out lung buds from early day 6 chick embryos in Howard's Ringer's. Cut just above shoulders and again just above hind limbs. Slice through body wall along one side and open embryo up. Remove internal organs. The lungs are located behind the heart and the bronchi lie along the esophagus. Remove extraneous tissue; lungs may separate into right and left lobes. Set Transwell 3452 (24 mm dia, 3.0 um pore size) tissue culture filter inserts in wells of a six well plate and underlay with 1.8 ml culture media each. Transfer explants to top of filter inserts. Explants of dorsal skin should be cultured with the mesenchyme side down. Remove any media on top of filters; explants should be cultured at the air:fluid interface, surrounded by a thin film of media that passes through the filter. Incubate at 37°C in a humidified incubator with 5% CO2.   Lung media: Ham's F12:DMEM (1:1), 1 ug/ml BSA, 10 ug/ml transferrin, 50 ug/ml gentamicin. References: Serra et al. (1994) Development 120:2153-2161. Souza et al. (1994) Development 120:2163-2174.

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