Gwen Stacy was a classmate of Peter Parker while they were undergraduates at Empire State University. Initially, Peter's problems as Spider-Man made him ignore her advances, and in return, she felt insulted by his aloofness. Gradually, however, a romance developed. Gwen, a science major, seemed to appreciate Peter's intellectual personality, different from that of jocks like Flash Thompson and preppies like Harry Osborn. She was Peter Parker's first true love.

On Earth-65, Gwen was the one bitten by a genetically-engineered spider and granted arachnid-like superpowers, and started a career as a crimefighter, dubbed by the media as "Spider-Woman." She was given a costume and a set of web-shooters by retired crimefighter Janet van Dyne. In this universe, Gwen is a free-spirited individual with artistic inclinations, traits that would put her at odds with the types of ethics her father tried to instill. She has an interest in music, hence her role as a drummer in the Mary Janes, and a love for corn dogs. Like her father, Gwen possesses a strong sense of justice and is a natural detective, both of which catered well to her decision to become a superhero after receiving her powers.