Carnival Corp's 704-passenger Adonia left port at about 4:24 pm (local time) yesterday.
The cruise comes after Cuba loosened its policy banning Cuban-born people from arriving to the country by sea. Cuban-born passengers were aboard.
Carnival says the Adonia will cruise every other week from Miami to Cuba.
Seventy-three-year-old passenger Rick Schneider told The Sun-Sentinel that he waited 60 years for the chance to make the journey. He bought a Cuban flag for the occasion, which he waved at protesters who opposed the cruises.