Avian influenza viruses (AIVs) are isolated routinely and propagated in specific pathogen free embryonated chicken eggs (ECE) and mammalian origin Madin-Darby Canine Kidney (MDCK) cell line. Continuous avian cell lines offer advantages for propagation of AIVs over MDCK cells because they maintain species specificity, and lower recurring costs compared to ECE. In this study, the characteristics of two avian fibroblast cell lines were evaluated, DF-1 (chicken-origin) and QT-6 (quail-origin), and their ability to support the growth of AIVs (n=19) belonging to nine different hemagglutinin subtypes from a variety of avian species. The replication efficiency of the AIVs in QT-6 and DF-1 cells was comparable to those in primary chicken embryo fibroblast (CEF) and MDCK cells. Receptor distribution analysis demonstrated high prevalence of SA alpha2,3-gal linked receptors in QT-6 and DF-1 cells which support a high growth of AIVs in these cell lines. Furthermore, the QT-6 and DF-1 cells supported high plaque-forming ability of representative highly pathogenic Eurasian H5N1 and H7N1 subtype AIVs. These two avian cell lines, especially QT-6 cells, also showed high transfection efficiency and could be useful for reverse genetics based rescue of AIVs. This study indicates that the DF-1 and QT-6 cell lines may be useful as a substitute for primary CEF and MDCK cells for AIV research in the areas of in vitro host range, molecular pathobiology and molecular genetics.