Avianflu could decimate Australian blackswans uq_news
]. Briefly, 17-day-old chicken and 21-day-old Pekin duck embryonated eggs were purchased from Darling Downs Hatchery . Primary aortic endothelial cells were cultured from the aortic arches of chicken and duck embryos as described previously using EGM-2MV medium with 10% FBS [305, 307]. All avian cells were cultured at 40 °C 5% CO2 unless otherwise stated.
The black swans used for cell isolation were sourced from Currumbin Wildlife Animal Hospital , Queensland, Australia, and Tauranga Harbor, Bay of Plenty, New Zealand . The animals were either euthanized due to a terminal illness or as part of a government-approved cull. Tibiotarsi and femurs were obtained from all birds. The bones were then partially opened in the middle to expose the bone marrow and then submerged in pre-chilled Dulbecco’s modified Eagle medium in T175 flasks.
Bone marrow was subsequently removed and resuspended in pre-chilled DMEM. A 15-ml suspension of DMEM containing bone marrow was then layered after filtering through a 40-μm sterile filter over 15ml of Lymphoprep® in a 50-ml SepMate® tube. The layer of cells at the DMEM-Lymphoprep interface was separated according to the SepMate manufacturer’s instructions. The separated cell layer was then resuspended in 1ml EGM-2MV media and enumerated.
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