- Pinocchio: A wooden boy puppet carved by Geppetto and turned into a living puppet by the Blue Fairy. Voiced by Seth Adkins.
- Patty: A new African American wooden girl puppet carved by Gladys and turned into a living puppet by the Blue Fairy returns. Voiced by Mallerie Rigsby.
- Geppetto: A kind and elderly woodcarver who created Pinocchio and wished for him to become a real boy. Voiced by Christian Rub.
- Gladys: A new kind and elderly woodcarver who creates Patty and wishes for her to become a real girl returns. Voiced by Jennifer Hudson.
- Figaro: A little kitty cat. Voiced by Kevin Crush.
- Cleo: A cute little goldfish. Voiced by Kelly Green.
- Jiminy Cricket: A cheerful and wise cricket who acts as Pinocchio's "conscience" and the partial narrator of the story. Voiced by Phil Criff Edwords.
- The Blue Fairy: Who brings Pinocchio to life and turns him into a real boy at the end of the film. Voiced by Rosalyn Landor.
- Angel 624 A Lilo & Stitch the Series the Coala Gilrs. Voiced by Tara Strong.
- Pater: A braid new England wooden boy puppet carved by Gordon and into a living puppet. Voiced by Haley Joey Osment.
- Gordon: A braid new kind and elderly woodcarver who creates Pete. Voiced by Rick Moranis.
- Stromboli: A large, sinister, Italian bearded puppet-maker who forces Pinocchio to perform onstage in order to make money. He speaks in an Italian accent and curses in Italian when he gets angry, though he is identified as a gypsy. Voiced by Chazz Pariminteri.
- Honest John & Gideon: A sly anthropomorphic red fox and the film's main antagonist who tricks Pinocchio twice in the film. Honest John's mute and crafty anthropomorphic feline sidekick. Voiced by Denis Leal and Billy Crystal.
- The Coachman: Owner and operator of Pleasure Island, who enjoys turning unruly boys into donkeys. Voice by Johnathan Harris.
- Lampwick: A naughty boy that Pinocchio befriends on his way to Pleasure Island; he is turned into a donkey on Pleasure Island. Voiced by Franky Daro.
- Monstro: The sperm whale that swallows Geppetto, Figaro, and Cleo during their search for Pinocchio. Pinocchio is later swallowed when Monstro is eating, and he and Geppetto reunite. Voiced by Don Nots'.
- When You Wish Upon a Star - Performed by: Spice Girls.
Disney Pinocchio with the Disney Guests Characters from the Episodes
Mickey Mouse and Friends - From: Pinocchio Meets Mickey Mouse and Friends
- Mickey Mouse
- Minnie Mouse
- Donald Duck
- Daisy Duck
- Clarabelle Cow
- Chip and Dale
Snow White and the Seven Dwarfs - From: Pinocchio and the Apple Trees
- Snow White
Tron - From: Pinocchio and Friends Meets Tron
The Little Mermaid - From: Pinocchio and the Little Mermaid
Aladdin - From: Pinocchio and the Arabian Nights
- Magic Carpet
The Princess and the Frog - From: Pinocchio in Bayou of New Orleans
The Haunted Mansion - From: Pinocchio at the Haunted Mansion
- Madame Leota
- Constance Hatchaway
- Erza the Skeleton
- Phineas the Traveler
- Gus the Prisoner
- The Singing Busts