Pouncey began attending the University of Florida early in 2007. His twin brother Mike would also join the Gators, playing defensive line his first season to help an injury-riddled defense. Pouncey saw action at right guard and center his freshman year. He started the season at right guard for the season opener against Western Kentucky, becoming only the seventh true freshman in team history to do so. In a week 6 loss to No. 6 LSU, he would have a grade of 86%. The next week he would have a grade of 85% while the Gators would upset the No. 8 Kentucky. On November 3, 2007 he would again get a champion grade of 83% in a win over Vanderbilt. He finished the season starting 11 games at right guard and playing in all 13. Rivals and College Football News selected him as a freshman first-team All-American and he would also be voted to the Freshman All-SEC team by the coaches and Sporting News.
Beginning in 2008, Pouncey became the full-time starting center for Florida next to his twin brother Mike at right guard. He compiled six games with a grade of 90% or better. The first came in week 3 against Tennessee, where he would have a grade of 93%. In week 6 he was recognized as the SEC Offensive Lineman of the Week for his performance against Arkansas. Pouncey would have a season-high grade of 97% against LSU and then 98% the next game against Kentucky. While playing South Carolina he would have a grade of 90, his last above that mark for the season. The Gators would go on to win the SEC Championship and then defeat Oklahoma to win the FedEx BCS National Championship.
In the season opener against Charleston Southern, he began 2009 with a grade of 98% and have two knockdowns in the game. The next week he would again post a grade of 98% in a victory over Troy. In the first SEC game of the year, Pouncey would play 61 snaps and receive a grade of 93% for the game. During the week 4 match-up against Kentucky, he would end with a grade of 90% and receive Champion Club Honors. On October 10, Florida traveled to Death Valley to face No. 4 LSU. He would have a season-high grade of 99% in the 13-3 victory over the Tigers. His 28th consecutive start would also be his last game at the Swamp. The Gators would go on to defeat Florida State 37-10. After going undefeated the entire regular season, the Gators would lose to No. 2 Alabama in the SEC Championship. last game would be the 2010 AllState Sugarbowl against No. 4 Cincinnati and he would finish his season with an average grade of 91.5% and 30 knockdowns. Pouncey won the Rimington Award, becoming Florida's first player to do so. He was also voted a first-team All-American by CBSsports.com, Sporting News, Football Weekly, and Walter Camp.
He finished his career with 39 starts in 41 games played. After his junior season he entered the NFL Draft.
His rookie year, he started in all 16 games played at the center position, and was selected to the Pro Bowl. Additionally, in accordance with his exceptional play as a center during his rookie year, he received 2 votes (out of 50) for the National Football League Rookie of the Year Award. In the AFC championship game during Pouncey's rookie year, Pouncey injured his ankle during the win and was not able to play in the Super Bowl. The Steelers would go on to lose the Super Bowl to the Green Bay Packers.
Pouncey played 14 games in 2011 then 15 games in 2012. Both those years, he was a Pro Bowl selection.
In Week 1 of the 2013 season, Pouncey tore his right ACL and MCL after Steelers guard David DeCastro accidentally dove into Pouncey's right leg while trying to assist Pouncey in blocking Tennessee Titans defensive tackle Sammie Lee Hill. He was subsequently placed on injured reserve, ending his season.
On June 12, 2014, the Pittsburgh Steelers signed Pouncey to a 5-year, $44 million contract extension. This extension made him, at that time, the highest paid center in the National Football League. After a successful recovery from knee surgery, Pouncey started all 16 games in 2014 and was selected to the Pro Bowl for the fourth time of his career.