A toy test subject and apostle of the Prototype characterized by blue fur and long limbs. He has multiple rows of sharp teeth. He can smash and tear down any obstacle. He is fast, cannot speak, and emits high-pitched screeches. He operates near the factory entrance.

A pink female spider toy test subject and apostle of the Prototype who can stretch her head, limbs, and torso. She pretends to be kind and fun on the surface, but is actually evil and insane. She operates in the Game Station.

A giant purple cat toy and apostle of the Prototype. He can emit red gas from his mouth to cause hallucinations. He follows the Prototype like a god and mercilessly kills anyone who interferes with him. He is quiet and operates in Playcare.

A kind and good-natured slime toy test subject. He can easily pass through narrow gaps and reconstruct his body by gathering pieces. He can also detach parts of his body, sharpen them, or harden them. He has three personalities: Jack, who is immature; Kevin, who is aggressive but good-natured; and Matthew, who is mature. Kevin's personality usually only appears when children are dying or in danger.

The ruler of the factory who has occupied it for 10 years based on a reign of terror and superior abilities. He is intelligent, possesses high speed, and can mimic the voices of others. He is obsessed with Poppy and continues experiments to create immortal life so that a situation where all other test subjects die, leaving only himself and Poppy lonely forever, never occurs.

A porcelain doll in the form of a young girl and the player's ally. She believes the Prototype is the root of all evil and wants the player to make him pay with his death. She is passive, weak, and dependent on others.

An ally of the Prototype and the manager of Playtime automation. He is cold and calm. He usually communicates by displaying a single large eye on a TV screen and uses spare units with TV heads during combat. He also commands the mass-produced Critters as his subordinates.

Guest's ally and a red bear mass-produced Critter.

A toy isolated in the manor who is bright and cheerful but possesses a somewhat twisted and mad personality. She intends to get into the Prototype's good graces and shows aggression if the player and Poppy's faction do not move according to her plans.

The protagonist of Poppy Playtime. They face off against the Prototype's apostles with excellent physical ability, reflexes, and quick thinking. They are a member of Poppy's faction.