Leila Hatami is the daughter of director Ali Hatami and is married to actor Ali Mosaffa. Having appeared in bit parts in some of her father's movies, Hatami's first lead film appearance was the title role in the 1996 film Leila, directed by Dariush Mehrjui. She received the Diploma of Honor for Best Actress from the 15th Fajr Film Festival.

In 2002 her role in The Deserted Station won the best actress award from the 26th Montreal World Film Festival. She has appeared in her husband's films as a director, Portrait of a Lady Far Away and The Last Step. In 2012 A Separation won the Academy Award for best foreign film in which Hatami played the leading female role.

In April 2014, she was announced as a member of the main competition jury at the 2014 Cannes Film Festival.

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