Voice cast
Shameik Moore as Miles Morales / Spider-Man: An intelligent and rebellious teenager of African-American and Puerto Rican descent, who is imbued with spider-like abilities after being bitten by a radioactive spider and eventually becomes the new Spider-Man. Producers Lord and Miller described the character as unique among Spider-Men because of his Brooklyn upbringing, his African-American and Puerto-Rican background, and the fact that his family is still alive, with that family dynamic being central to the film's story.
Jake Johnson as Peter B. Parker / Spider-Man: Miles' reluctant mentor, a disheveled, jaded and brown-haired 38-year-old counterpart of Spider-Man from another dimension. The character is intended to be an amalgamation of all pop culture Spider-Man adaptations and interpretations. Lord and Miller envisioned him to be like Mr. Miyagi from The Karate Kid franchise, if "Mr. Miyagi doesn't know anything", which they thought was a "really neat color to put onto Peter that we hadn't seen before".
Hailee Steinfeld as Gwen Stacy / Spider-Gwen / Spider-Woman: A dimensionally-displaced counterpart of Gwen Stacy with spider-like abilities, who takes up the alias of "Gwanda" while at Miles' school. In onscreen comic books while explaining her origin story, the character is referred to as "Spider-Gwen". Her relationship with Miles is depicted as a growing friendship, while he displays outward romantic feelings on his end. This decision was made to avoid relegating Gwen to being a love interest and to help her to be defined by her capability as a hero.
Mahershala Ali as Aaron Davis / The Prowler: Miles' uncle, who moonlights as a masked enforcer for Kingpin while keeping it a secret from his family. The character is rebellious, causing him to be looked at poorly by Jefferson.
Brian Tyree Henry as Jefferson Davis: Miles' African-American and police officer father, who disapproves of Spider-Man's vigilante actions. At the age of thirty-five, Henry had said he was too young to portray a father of a teenager, but agreed to the role after learning that Miles Morales was the only black and Latino Spider-Man.
Lily Tomlin as May Parker: Peter's aunt, who is deceased in Peter B. Parker's universe and provides refuge for the other Spider-People in Miles' universe.
Luna Lauren Vélez as Rio Morales: Miles's Puerto Rican and nurse mother.
Zoë Kravitz as Mary Jane "MJ" Parker: Peter Parker's wife in Miles's universe and Peter B. Parker's ex-wife in his universe.
John Mulaney as Peter Porker / Spider-Ham: A talking animal-based Spider-Man from a cartoon-based anthropomorphic universe, who was once a spider, bitten by a radioactive pig.
Kimiko Glenn as Peni Parker / SP//dr: A young Japanese-American girl from an alternative and anime-based universe who copilots her late father's biomechanical suit with a radioactive spider that she shares a telepathic link with. The filmmakers initially considered using Silk as their Asian-American Spider-Man, but eventually settled on Peni because of her more distinctive power set compared to the other Spider-People. The character's designs went through a few iterations as her initial design was particularly "iffy". Production designer Justin Thompson considered Lord and Miller's desire to use an anime design and came up with the idea to portray her in an art style similar to Sailor Moon.
Nicolas Cage as Peter Parker / Spider-Man Noir: A Spider-Man from a monochromatic universe set in the 1930s. Cage based the character on the films of Humphrey Bogart and voices of actors from that era such as James Cagney and Edward G. Robinson.
Kathryn Hahn as Doctor Olivia Octavius / Doctor Octopus: The head scientist and CEO of Alchemax and a scientific adviser to Kingpin. The character uses highly advanced tentacles and is featured in much of the marketing for Alchemax, even being seen in a documentary shown at Miles' school.
Liev Schreiber as Wilson Fisk / Kingpin: A crime lord and the benefactor of Alchemax in Miles' dimension. The character is heavily responsible for the creation of the collider and is attempting to bring an alternate version of his deceased family into his dimension.
In addition to the six main versions of Spider-Man present in the film, Chris Pine is featured as the version of Peter Parker's Spider-Man native to Miles' dimension. Though the character offers to mentor Miles, he is murdered by Kingpin following the collider's activation before he can do so. This iteration of Parker was intended to be "as competent a Spider-Man as possible" and combines elements from previous Spider-Man portrayals, but with slight differences to indicate that he is from a different universe than the others. According to Parker's eulogy, he was a graduate student and married to Mary Jane Parker.
Additional voices for the film include: Lake Bell as Vanessa Fisk, Kingpin's wife, Jorma Taccone as Norman Osborn / Green Goblin, a genetically-enhanced mercenary working for Kingpin, Marvin "Krondon" Jones III as Tombstone, Kingpin's silent and heavily-armed bodyguard, Joaquín Cosío as Scorpion, a Mexican cyborg with scorpion-like prosthetics and Post Malone (who contributed to the film's soundtrack) as a bystander in Brooklyn. An archival recording of Cliff Robertson from Spider-Man 2 (2004) was used for a flashback scene involving the character Ben Parker. Spider-Man co-creator Stan Lee appears in a posthumous cameo, as a character named Stan who sells a Spider-Man costume to Morales. Lord and Miller said it was important to give Lee a bigger moment in the film rather than just a passing cameo, because he was "so integral to the spirit of this movie", and the role was "extra meaningful" following Lee's death in November 2018. Lee's character also has several brief "Easter egg" cameos throughout the film, such as when he walks over Miles and Peter B. when they are lying on the streets of New York City. A young Matt Murdock from Gwen's dimension appears in an "Easter egg" cameo during Miles's final confrontation with Kingpin as the latter's alternate universe adopted son.
Cameos during the film's post-credits include Oscar Isaac as Miguel O'Hara, a Spider-Man from the Marvel 2099 imprint, Greta Lee as O'Hara's AI assistant Lyla (respectively credited as "Interesting Person #1" and "Interesting Person #2") and Jorma Taccone as the Spider-Man from the 1967 animated series (replacing Paul Soles, with the character being credited as "Last Dude"). Donald Glover appears on a background TV screen, which is displaying a scene from the episode "Anthropology 101" of Community in which Glover's character, Troy Barnes, wears Spider-Man pajamas. This scene, a reference to an unsuccessful Twitter campaign suggesting Glover be cast in the lead role of The Amazing Spider-Man (2012), was cited by Miles Morales's creator, Brian Michael Bendis, as influencing the design and portrayal of the character. Glover had previously voiced a version of Miles Morales in the Ultimate Spider-Man animated series, and played Aaron Davis in the Marvel Cinematic Universe (MCU) film Spider-Man: Homecoming (2017).
Miles Morales's best friend and roommate Ganke Lee appears. The character originally had a bigger role, but was rewritten due to Spider-Man: Homecoming having the similar character Ned Leeds. Pixar animator Peter Sohn was cast as Ganke before the character's dialogue was cut from the final film; Sohn would later voice the character in the sequel, Spider-Man: Across the Spider-Verse (2023).