Harry, 31, slipped into the park late at night after attending the U.S. vs. U.K. sitting volleyball match at the Invictus Games, held at the nearby ESPN World Wide of Sports Complex. Harry visited Splash Mountain, a log flume ride, Space Mountain and other Disney attractions.
He spent a little less than one hour at the park.
A Kensington Palace spokesman confirmed the visit, saying, "He went into the park last night after a full day of watching great Invictus competition."
Prince Harry told "Good Morning America" co-anchor Robin Roberts in March that the trip holds "very, very happy memories" for him.
"It's one of my very, very happy memories, of going to Disney World with my mum," Harry said. “I went on ‘Space Mountain’ 14 times. I was like, 'This is absolutely fantastic. This is the best thing ever.'"
Earlier on Tuesday, Prince Harry lost a $20 bet to U.S. veteran Israel del Toro of Colorado Springs, Colorado, during Team USA's volleyball final against Team Great Britain.
The two challenged each other by each betting on their respective nation's victory. The United States took top honors, winning the gold medal.