The reproductive biology of the invasive blue crab Callinectes sapidus was studied in the Thermaikos Gulf (Northwest Aegean Sea, Eastern Mediterranean). In the two-year survey, 5698 (2897♂/2801♀) crabs were caught with the use of fyke nets. Total sex ratio (♂/♀) reached equality (1.03:1). The female blue crab exhibited a protracted reproductive period. Mature and ovigerous females exhibit short migratory movements from estuarine and inshore waters, where the population mostly congregates (0–3 m), and move to slightly deeper waters (1–3 m) up to 9 m for spawning. A total of 340 ovigerous females were caught. Their number varied both spatially and temporally; they were observed for a 7-month period (April to October) with a clear peak in July–August and at a 3 m depth gradient corresponding to ≈60% of the total number of ovigerous females caught in both years. Size at first sexual maturity (CW50) was estimated at 113.1 mm CW. Average fecundity was ≈790,000 eggs. Experimental trawling showed that inshore waters (<1 m) in the estuaries serve as nursery areas for juveniles. Defining the spatiotemporal and bathymetrical distribution of ovigerous females in any invaded coastal habitat could be considered key information for the implementation of a management policy for the species.