Usain Bolt played 20 minutes for Norwegian team Stromsgodset in a friendly against Norway U19s on Tuesday.
The world's fastest man, who retired from athletics last year, has openly discussed a desire to play professional football, stating in 2016 that his dream was to play for Manchester United.
The Jamaican will play at Old Trafford at Soccer Aid this weekend.
Bolt wore the number 9.58 for Stromsgodset - the time of his 100-metre world record set in 2009 - as the team lost 1-0.
The 31-year-old was a late substitute and, reportedly, was given the best chance of the game to equalise.
Former Borussia Dortmund head coach Peter Stoger said earlier this year that Bolt has "a lot of work to do" to become a top-level footballer, after he trained with the Bundesliga side.
"The physique he needs for his other sport is completely different from what he needs for football, but it was really fun for us. It was a pleasure for us to meet a guy like him and to work with him," Stoger said.