The azulgrana team, which reached the semifinals last season, will face the first match at home and the rojiblanco will do so as a visitor.
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The French Olympique, defending champion and seven-time champion, will play their continuity against the Danish Brondby, a team that they left out in the 2011-12 campaign in the quarter-finals and that have been in the competition for 18 seasons.
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Pernille Harder: “I signed for Chelsea because they want to win everything”
Both Lyon and Barcelona have won all ten of their round of 16 matches in the last five seasons.
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The French are the first team to have won seven titles and five of them consecutively after 2020, in addition to being the only team to have reached the round of 16 in the 12 seasons with the current format of the competition.
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German Wolfsburg, the last runner-up, was paired with Norwegian LSK Kvinner and Paris Saint Germain, who also played in the semi-finals and was eliminated by Lyon, will face Czech Sparta Prague.
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