Variability between Aedes aegypti populations in north Queensland, Australia, has the potential to impact the successful implementation of new population replacement mosquito releases for dengue control. Four Ae. aegypti colonies originating from different locations (Cairns, Mareeba, Innisfail, and Charters Towers), along with one F1 field-derived population from Cairns, were inter-crossed to determine any incompatibilities in copulation, insemination, and production of viable offspring. Greater copulation and insemination rates were observed when males recently introduced from the wild (‘Cairns-Wild’ population) were mated with long-term laboratory females. Egg viability rates for all crosses ranged from 90.2–98.2%, with no significant differences observed between crosses. Greater egg production was seen in some populations, and when corrected for wing-length, egg production was greatest in a Mareeba × Innisfail cross (19.55 eggs/mm wing length) and lowest for the Charters Towers intra-population cross (14.35 eggs/mm). Additionally, behavioral differences were observed between laboratory and wild mosquitoes from the Cairns location, suggesting possible laboratory conditioning. Finally, despite controlled larval rearing conditions, size differences between populations existed with Charters Towers mosquitoes consistently smaller than the other populations. The spread of genes or bacterial symbionts between these populations is unlikely to be hindered by pre-existing reproductive barriers.