Barbie makes her first brief appearance as a character in Toy Story 2, where she is among the many Barbie dolls guiding Andy's toys on their quest to rescue Woody. Her character is further developed in Toy Story 3, where she resides at Sunnyside Daycare alongside Ken. Initially, Barbie and Ken clash, but they eventually reconcile and become a couple, collaborating to enhance the daycare experience for all the toys. She makes a brief appearance during a flashback scene when Woody, Buzz, Jessie, and Slinky are trying to rescue R.C. from outside in a thunderstorm in Toy Story 3.

Mattel had been hesitant to allow the use of Barbie's image in the movie due to concerns about potential negative impacts on the doll's reputation. They feared that the film's portrayal might not resonate with audiences and could undermine Barbie's longstanding appeal. Additionally, Mattel aimed to preserve Barbie's neutrality, enabling buyers to envision her in various ways rather than being confined to a specific movie depiction. As a result of these concerns, the role originally intended for Barbie in the first film was ultimately given to Bo Peep.