Bats are natural reservoir hosts for several viruses of public health concern, many of which can cause severe disease in humans and other susceptible hosts but cause no clinical disease in bats. The Australian black flying fox (Pteropus alecto) is a reservoir for Hendra, Menangle, and Australian bat lyssavirus, among others. In addition, the pteropine orthoreoviruses (PRVs) are carried by a variety of bat species in Australia and Southeast Asia and are considered emerging zoonotic viruses capable of causing severe respiratory disease in humans. Understanding how bats control viral infections could help prevent spillover to other species and uncover novel therapeutic targets. To provide a more physiologically relevant model to study the host response of bats to respiratory viruses, we isolated airway epithelial cells from P. alecto tracheobronchial tissue and differentiated them at the air-liquid interface to form an in vitro pseudostratified epithelium. The well-differentiated P. alecto cells were characterised and their responses to infection with a PRV (Nelson Bay virus) or following challenge with the dsRNA mimic, poly(I:C) were examined in comparison to human cells. The well-differentiated P. alecto cells formed a mucociliary phenotype, characterised by mucus secretion and beating cilia on the surface of ciliated cells. Histology and immunostaining confirmed the formation of a pseudostratified epithelium consisting of basal, goblet, club, and ciliated cells. Stimulation of the differentiated P. alecto airway epithelium with Poly(I:C), confirmed that the P. alecto cells could mount an immune response. Additionally, infection of differentiated bat and human airway epithelia with the Nelson Bay virus exhibited diverse Type-I and -III interferon responses despite both being resistant to the virus strain, although at different levels. In this study, we have developed a bat airway epithelial model from a known reservoir native species, demonstrating its physiological relevance for studying host-pathogen interactions and the innate immune features.