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Born Gloria Frances Stewart in Santa Monica, California, she changed the spelling of her name when she commenced her acting career.

After acting in college and in other amateur productions, Stuart was signed to a contract by Universal Studios in 1932. She was also selected as one of the thirteen WAMPAS Baby Stars of 1932. As a glamorous blonde, she was quickly cast in a variety of films and became a favourite of director James Whale, appearing in his The Old Dark House (1932), The Invisible Man (1933) and The Kiss Before the Mirror (1933). Her career with Universal Studios failed to gain momentum and she moved to 20th Century Fox. By the end of the decade she had starred in more than forty films, including Roman Scandals (1933) and Rebecca of Sunnybrook Farm (1938) but had not become a major star.

She made a few films during the 1940s but had become more interested in working with the Screen Actors Guild, and in developing her talents as a painter. As an artist she became well respected with her work being exhibited throughout the USA and Europe. After a thirty year break from acting, she appeared in the 1975 television movie The Legend of Lizzie Borden and over the next few years appeared regularly on television. She made her first cinema appearance in almost forty years when she appeared in My Favourite Year in 1982.

In old age, she achieved a level of celebrity she had never experienced during her years as a Hollywood contract player, when cast in Titanic (1997). As the centenarian Rose she received an Best Supporting Actress Academy Award nomination, becoming the oldest person to receive an acting nomination.

Stuart found herself in demand after this and was constantly employed, with her most recent role being in a Murder, She Wrote TV movie in 2001.