[2][3] Location of South African teams Sixteen teams qualified for the 2024–25 EPCR Challenge Cup from Premiership Rugby, the Top 14 and the United Rugby Championship as a direct result of their domestic league performance having not qualified for the Champions Cup.

Two further sides received invitations, South Africa's Cheetahs and Georgia's Black Lion.

The knockout stage begins with a single-leg round of 16 matches, consisting of the top 4 teams from each pool, and the teams ranked 5th in each pool of the 2024–25 European Rugby Champions Cup.

The round of 16 follows a pre-determined format, while the quarter-finals and semi-finals guarantee home advantage to the higher-ranked team.

Note: Flags to the left of player names indicate national team as has been defined under World Rugby eligibility rules, or primary nationality for players who have not yet earned international senior caps.