Hamster & Gretel features LGBTQ+ characters, including Carolina Grant-Gomez who has two moms and same-sex crushes, and Professor Exclamation who is non-binary. Their identities are normalized and presented respectfully, contributing to an affirming portrayal without being central to the main narrative or a source of conflict.

Hamster & Gretel features Laurette, a recurring transgender character voiced by a trans woman. Her identity is revealed in a matter-of-fact, affirming way, and she is consistently portrayed with dignity, competence, and respect. The show integrates her seamlessly into the narrative, avoiding stereotypes or negative framing, thus presenting a supportive and validating depiction of a transsexual individual.

The show features Gretel Grant-Gomez, a female superhero with superpowers, who frequently engages in and wins close-quarters physical fights against various male villains. Her victories are achieved through direct physical confrontation and the application of her superhuman abilities.