Shameik Moore as Miles Morales (voice)

Hailee Steinfeld as Gwen Stacy (voice)

Brian Tyree Henry as Jeff Morales (voice)

Luna Lauren Velez as Rio Morales (voice)

Oscar Isaac as Miguel O'Hara (voice)

Jake Johnson as Peter B. Parker (voice)

Jason Schwartzman as Spot (voice)

Issa Rae as Jessica Drew (voice)

Daniel Kaluuya as Hobie Brown (voice)

Nicolas Cage as Spider-Man Noir (voice)

John Mulaney as Spider-Ham (voice)

Kimiko Glenn as Peni Parker (voice)

Amandla Stenberg as Margo Kess (voice)

Karan Soni as Pavitr Prabhakar (voice)

Mahershala Ali as Uncle Aaron (voice)

Jharrel Jerome as Miles G. Morales (voice)

Greta Lee as LYLA (voice)

Andy Samberg as Ben Reilly (voice)

Shea Whigham as George Stacy (voice)