In The Ballad of Gay Tony, Luis Fernando Lopez must drive his friend Al Di Napoli to a drug deal with Oscar Gomez and another unknown accomplice at the university. During the mission, the deal is ambushed by police, and winds up with the four escaping.
- The university was likely named after Amerigo Vespucci, the explorer whom the continents of North and South America were named after. This is again to mirror Columbia University, named after Christopher Columbus, an explorer of North and South America.
- Ruby St. runs through the University.
- Niko Bellic lies when he states on his resumé to Goldberg, Ligner & Shyster that he attended the School of Law at Vespucci University.
- Brian Meech graduated from Vespucci University.
- The Higgins Helitours' pilot describes Vespucci's library as the place "where they get their long hair stuck in their fancy books."
- Molly Schultz has a law degree from Vespucci University.
- Georgina Cheng completed her masters degree at Vespucci University.