Penicillin streptomycin amphotericin b suspension 100 antibiotic antimycotic solution

Before poly:IC exposure, we cut the formed organoid with ophthalmologic scissors to approximately one-fourth the size under a stereoscopic microscope due to the reduction of variety of sizes. The organoids were exposed to 5 μg/mL or 50 μg/mL poly I:C (polyinosinic-polycytidylic acid sodium salt) (4287/10; R&D Systems, Minneapolis, MN) for 6 h. We randomly chose nine-pieces from each dish (exposure of 0 μg/mL, 5 μg/mL, 50 μg/mL poly:IC) from a total of 36 pieces of organoids. During preculture and poly I:C exposure, the cells were cultured in DMEM. The DMEM (043–30085; FUJIFILM Wako Pure Chemical Industries) medium contained 10% FBS (SH30396.03; Cytiva) and 1% Penicillin-Streptomycin-Amphotericin B Suspension (×100) (Antibiotic-Antimycotic Solution) (161–23181; FUJIFILM Wako Pure Chemical Industries). Cells were cultured at 37°C in 5% CO₂.

We used KAv-1 medium to obtain and culture chicken, Okinawa rail, domestic duck, and whooper swan fibroblasts. KAv‐1 is based on α‐MEM and contains 5% FBS (SH30396.03; Cytiva, Marlborough, MA, USA), 5% chicken serum (16110082; Thermo Fisher, Waltham, MA, USA), 0.1% D-glucose (041–00595; FUJIFILM Wako Pure Chemical Industries, Osaka, Japan), 0.4 mM calcium chloride, 10 mM EPPS (348–03192; FUJIFILM Wako Pure Chemical Industries), 0.11% sodium carbonate (199–01585; FUJIFILM Wako Pure Chemical Industries), 55 μM 2-mercaptoethanol (21985–023; Thermo Fisher, Waltham, MA, USA), and 1% penicillin-streptomycin-amphotericin B suspension (×100) (Antibiotic-Antimycotic Solution) (161–23181; FUJIFILM Wako Pure Chemical Industries). Cells were cultured at 37°C in 5% CO₂.