Born in 1958, in Hitchen, Hertfordshire, England, his father, Trevor Bartlett, was a Physical Education teacher. Neil Bartlett featured him in his play Night After Night. In 1987 Neil Bartlett formed the Gloria theatre company. He devised and performed in Sarrasine, a work of musical theatre from Honoré de Balzac's (1799-1850) story.

He also wrote and performed in A Vision of Love Revealed in Sleep, which was inspired by the life of the Pre-Raphaelite artist Simeon Solomon who continued to paint and love boys despite poverty and being arrested for having sex with a man. The play draws in Simeon Solomon's 1871 prose poem with the same title. Neil Bartlett performed the solo show in a warehouse off Tooley Street, south London. The production dramatises a young man's journey from fear to revelation.

In 1991 Neil Bartlett's translation of Racine's French classic play Bérénice was staged at the National Theatre, and his adaptation of Molière's School for Wives which was produced at the Derby Playhouse. In 1994 he became the artistic director of the Lyric Theatre, Hammersmith, London. His first production The Picture of Dorian Gray was highly acclaimed. In 1995 he staged a translation of Jean Genet's play Splendids, with Julian Clary.