Dottie "Doc" McStuffins (voiced by Kiara Muhammad) - The main character of the series. Doc is a six-year-old girl who likes to fix toys, dolls and stuffed animals. She also wants to be a doctor like her mother, one day. She is a girl with short dark brown hair tied in pigtails with a fashionable headband in the middle of her head. She usually wears a lab coat over a purple-and-white striped T-shirt and pink-colored skirt with polka-dotleggings on her legs and violet ankle socks with pink sneakers. The only other clothes she has been shown in is a large Indiana Jones type sunshade hat added to her usual attire or a BucketHat in a camping sequence. As well as a straw bonnet during a tea party sequence, a pink polka-dotted dolphin tankini in a pool/water park sequence, a SilkScarf, white ankletsocks, sunglasses and a pair of high heel shoes while pretending to be a pop star, a cowboy costume in a Halloween episode and a winter outfit in a Christmas episode and a pair of pajamas with a pair of bunny slippers in a bedtime sequence. In Season 2, Doc wears a School uniform consisting of a white short sleeve PoloShirt ribbon tie, plaid skirt, white knee socks and a pair of Mary Janeshoes and in the Christmas episode wore a snow outfit with Ugg type boots. Her name is also used for the series title.
Lambie (voiced by Lara Jill Miller) - A stuffed lamb who is one of Doc's best friends. She is very sweet, likes to hug people and is a gifted dancer who wears a pink bow with her matching tutu. Her other friends are Stuffy, Hallie and Chilly. She has a crush on Stuffy because she gives him the most cuddles.
Stuffy (voiced by Robbie Rist) - A stuffed blue dragon who tries to be the bravest dragon of all but does not succeed at that every time especially with spiders which he has a fear of. He is also clumsy. He is one of Doc's friends and his other friends are Hallie, Lambie and Chilly. His design is partially inspired by Figment from the Epcot ride Journey Into Imagination.
Chilly (voiced by Jess Harnell) - A stuffed snowman who worries a lot, and does not always seem sure that he isn't a real snowman. He is sweet and one of Doc's fearful friends. He is also friends with Lambie, Stuffy and Hallie. Chilly wears a black bowler hat & a blue scarf.
Hallie (voiced by Loretta Devine) - Hallie is a stuffed hippo and a nurse who is Doc's assistant. She is friendly and treats Doc's patients with respect and good care. She is good friends with Doc and the other stuffed animals. She also has a Radar O'Reilly-like intuition as to what Doc needs during an exam, which she calls a "Hippo Hunch." She may be a tribute to Henrietta Hippo from the classic 1970's children's show The New Zoo Revue. She wears a Candy striper nurse's outfit.
Squeakers (voiced by Ian Anthony Dale) - A purple squeaky toy that looks like a blowfish and can go in the water like Hermie, Marvin, Surfer Girl, Melinda the Mermaid and Lula.
Donny McStuffins (voiced by Jaden Betts) - Donny is Doc's four-year-old brother who usually spends most of his time playing with his toy cars and his friends. He owns the toy knight action figure named Sir Kirby, who appears in the episodes Knight Time and The Dark Knight.
Dr. McStuffins / Mom (voiced by Kimberly Brooks) - Dr. McStuffins is the mother of Doc and Donny and the wife of Mr. McStuffins. She is a doctor who works at a hospital. She expects Doc to become a doctor, like herself.
Mr. McStuffins / Dad (voiced by Gary Anthony Williams) - Mostly a stay-at-home father but indicated in "The New Girl" to have a part-time job who usually cooks dinner while his wife is at work. He takes care of Doc and Donny, his children.
Note: Doc McStuffins is the only human character in the show that can talk to toys.
Emmie (voiced by Kylee Anderson) - The girl next door, Doc's soccer teammate, and best friend.
Katherine -Doc and Donny's babysitter neighbor who once owned Dress-Up Daisy
Jacob -The boy in the park, who owns Super Stuntman Steve.
Toys in the Clinic
Aurora (voiced by Laraine Newman) - A telescope belonging to Henry, who speaks in 60's hippie lingo and walks on her tripod. The night she came out of the box, her vision was strangely blurred, and Doc led a search of Henry's yard to find Aurora's missing eyepiece.
Awesome Guy (voiced by David Boat) - A toy superhero that belongs to Donny, he had to rescue Lambie from a tree, but there was a problem with his legs, Doc wanted to give him a checkup, but he didn't want to get one, and later Stuffy had an idea to sneak him in, Doc found out the problem, his legs were put on the wrong way, so when Doc fixed them he was back in action.
Bella (voiced by Julianne Buescher) - A ballerina like Lambie, only she's human and wears a blue outfit and speaks in a Russian accent. She accidentally cracked her leg after a leap, and had to have it put in a cast, but she found lots to do whilst she waited for it to heal.
Ben & Anna (voiced by Jeffrey Nicholas Brown and Meghan Strange) - Two stuffed monkeys that stick to each other, and they belong to Alma. Ben had to be fixed when his Velcrosticker ripped off and he got separated from Anna overnight.
Big Jack (voiced by Ty Burrell) - A toy box with a joker popping out when the handle turns.
Boomer (voiced by Dave B. Mitchell) - The soccer ball that belongs to Emmie. He was flat due to a leak, but was scared of the needle in the air pump Doc had to use.
Boppy (voiced by James Arnold Taylor) - The blue inflatable punching bag shaped like a dog, who usually can't be knocked down. He finds himself losing air after a rosebush popped him, and was taken in by Doc for patching and reinflation (This scene is used as part of each show's opening).
Bubble Monkey (voiced by Hynden Walch) - A monkey that blows bubbles and belongs to Emmie and Alma. She stopped blowing bubbles when Alma accidentally filled her with paste instead of bubble soap.
Buddy (voiced by Jess Harnell) - A dump truck toy that belongs to Donny. Buddy is Riggo's best friend. (Based on the Buddy-L line of toy trucks.
Choo-Choo Train- A wooden toy train who is used either as a ambulance to move the smaller injured toys or as a get away vehicle for the Wicked King.
Dress-Up Daisy -A fashion Doll that once belong to Katherine when she was Doc's age and was presumed lost after a gust of wind took her away while wearing a parachutist outfit
Frida (voiced by Sutton Foster) - A cute toy fairy kite who thinks she's a real, magically fairy.
Gabbi (voiced by Lacey Chabert) - A toy giraffe that Doc won at the arcade. She had a ripped leg, so when Doc fixed it, she decided to take her home.
Glo-Bo (voiced by Jim Belushi) - A toy monster with six arms, he is a friendly toy monster with a joyful personality. Neglected for several weeks when Donny got a new train, Glo-Bo consulted Doc about his lost ability to glow in the dark. He also inadvertently spread a rash (which was really blue paint) to lots of toys, including Chilly.
Gustave (voiced by Stephen Stanton) - A green plastic alligator with a Cajun accent and appetite for marbles. He is one of four mechanical players in Donny's game Gulpy-Gulpy Gators (a spoof of Hungry Hungry Hippos). Doc had to empty his stomach after Donny added extra marbles to the game and Gustave wound up overstuffed.
Hermie (voiced by Ari Rubin) - A toy crab that Doc, Squeakers, and Marvin can play with in the water. He once lost an arm after a rough run-in with Rudi the pup.
Kiko (voiced by Janice Kawaye) - A Japanese Anime / Manga doll Doc got from her Grandma. Her problem was having frail legs and being stuck in her box from her long flight from Japan but Doc McStuffins helped her exercise.
Lenny (voiced by Jeff Fischer) - A toy firetruck that belongs to Donny, he is also known as Engine 9. Donny put him in the pile of toys that he doesn't play with anymore because he didn't squirt out any water from his hose. Doc refilled him with water and returned himself to Donny.
Lelani - A solar powered hula girl
Little Jack (voiced by Nolan Gould) - The son of Big Jack. He had trouble popping out from his box. He was scared to get a check-up, but then wasn't scary. Then Doc found out that his clothes were stuck between the cogs of the music box.
Loud Louie (voiced by Georgina Cordova) - A toy cellphone that Doc used to play with when she was little, he was loud because he was super excited, so he had to learn to use his inside-voice.
Lula (voiced by Grey DeLisle) - A sponge toy that looks like a Beluga whale, when Lula is placed in water it takes a couple hours for her to grow, in the end she has grown to fit in the fish tank in Doc's clinic.
Marvin A squirt toy shaped like a duck who lost its squeaker at the water park.
Melinda the Mermaid (voiced by Dharbi Jens) - A plastic toy mermaid that was found at the kiddie pool, she had trouble swimming because her winder-upper was stuck and she didn't take swimming lessons, so when she took them, she can swim now.
Millie the Microphone (voiced by Lisa Loeb) - Emmie's toy CD boombox with a face. Millie kept repeating words when she spoke because her disc was dirty inside her.
Moo Moo (voiced by Colette Whitaker)- A stuffed cow belonging to Alma, who drags out her long u's when she speaks, so it sounds like she's actually mooing. Once she didn't want to go back to Alma after being left out in the rain.
Mr. Chomp (voiced by Brad Abrell) - The plastic toy shark that belongs to Donny, he's very kindhearted and doesn't bite like a real shark.
Niles (voiced by Charlie Schlatter) - Donny's toy crane, he has a bandage on his crane because his crane was unfixable, but Niles didn't want his bandage changed because he was nervous about getting it removed, later he was brave enough to get his old bandage removed slowly and quickly.
Officer Pete (voiced by Michael Gough (voice actor))- Donny's metal toy police car who takes his job very seriously and overheated while pusuring The Wicked King in a very hot Summer day.
Penny Possum (voiced by Audrey Wasilewski) - Penny is the mother of three possum children, Pip, Flip, & Trip. The children went missing when one of Penny's velcro patches ripped loose.
Pickles the Bunny (voiced by Colleen O'Shaughnessey) - She's a stuffed bunny that wears bows on her ears and has a heart on her chest, she was in a box at Alma's yard sale and left because she thought that she wasn't loved anymore, later Doc finds out that Pickles triangle nose is missing and replaces it with a button, soon Doc returns Pickles to Alma and puts her in her room.
Professor Hootsburgh aka "Hootsie" (voiced by Laraine Newman) - A stuffed owl with a very smart brain. Doc got her from her older cousin Tisha.
Ricardo Racecar (voiced by Ian Gomez) - Donny's Italian toy race car who lost energy because Donny raced him and didn't let him recharge, but when Doc's dad put him onto charge, he was soon better.
Riggo (voiced by Dennis Farina) - A grader toy that belongs to Will, Riggo is Buddy's best friend, his scooper was stuck because there was sand on the side of his scoop, jamming his gears. But Doc didn't figure this out until she suffered a little sand in her own eye.
Robot Ray (voiced by Dee Bradley Baker) - Donny's battery-operated robot who considers himself a rescuer like Ronda, but once he finds out that he can't get wet in the episode All Washed Up. Ray is equipped with long-range vision and a white cord he shoots out with his right hand, extending his reach. He was also at Hallie's birthday and went camping with Doc, Donny and their Dad in the episode "Out in the Wild". He broke his sticky launcher, but Doc used first-aid to fix his arm till the end of the campout.
Rescue Ronda (voiced by Camryn Manheim) - The remote-control helicopter that belongs to Luca. Her job and passion is rescue missions, and has ended up in Doc's clinic more than once with damage taken in the line of duty. If Rescue Ronda can't perform a rescue herself, she'll direct her friends on how to help. (Her name is a possible spoof of The Beach Boys hit song, "Help Me, Ronda.")
Sebastian (voiced by Dee Bradley Baker) - He's a ghost that lives in a toy pumpkin, he was scared of Halloween, but Doc explains to him that it's all pretend, so he decides to go back to his job.
Sir Kirby (voiced by Rob Paulsen), - The armoured knight action figure belongs to Donny, who often plays the hero in Doc's game "Save Princess Lambie from Stuffy", but not without taking some damage.
Star Blazer Zero (voiced by John Michael Higgins) - A toy alien who pilots a rocket ship, his catchphrase is "Kazowiee", his arms and legs fell off because he didn't wear his seat belt.
Super Stuntman Steve-A extreme stunt motorcyclist action figure who belongs to Jacob.
Surfer Girl (voiced by Kimberly Brooks) - A teenage girl with her feet attached to a surfboard.
Susie Sunshine (voiced by Amber Hood) - A plastic toy doll that wears a pink dress and pink shoes. She was cranky because her eyes were stuck open because a little pineapple ice pop was stuck on her eyebrows.
Teddy B. (voiced by Jason Marsden) - Donny's teddy bear who was reunited but caused Donny to sneeze due to an excess of dust on his fur for being laying around and not being played with for ages.
Tremaine (voiced by Ari Rubin) - A toy truck designed by Donny, he is very dynamic, he needed to stand still because he had to get his stripes repainted.
Val (voiced by Jennifer Hale) - A heart shape stuffed toy given by Doc's parents for Valentine's Day and causes a rift between her and Lambie.
Walter & Gracie (voiced by Tom Kenny and Grey DeLisle) - Two walkie talkies that are both different genders, Walter lost his antenna and Gracie was lost in the strawberry field.
Wicked King (voiced by Jess Harnell) - He sometimes acts like an evil toy when he's involved in playtime, he resembles an old Fisher Price's Little People toy because he has no hands or arms. He's also known as Percy, his real name.
Xyla (voiced by Tiffany Thornton) - A toy xylophone that looks like a ladybug and belongs to Alma, one of her keys was loose because one of the screws was loose, and when the key fell into the sink, Doc replaced it with a new one.