Spotify quickly pulled the ad after complaints, admitting it could be seen as "culturally insensitive".
"We made a creative decision to feature Justin Bieber in our ad because we wanted to celebrate 'Despacito' as key cultural moment when music genres crossover," a spokesperson told the New York Daily News.
"We realised that this could be seen as culturally insensitive so we have pulled those ads."
The incident comes after Bieber was filmed performing Despicato while struggling to sing the song's Spanish words.
"I don't know the words, so I say poquito/ I don't know the words, so I say Dorito," he was reported as singing.