The streaming company suspended production earlier this week of the upcoming sixth and final season of the political intrigue drama, in which Spacey plays U.S. president Frank Underwood.
"Netflix will not be involved with any further production of 'House of Cards' that includes Kevin Spacey," the company said in a statement late on Friday.
Spacey's publicist cut ties with him on Thursday and it was not known who currently represents him.
His departure may not mean the end of the show, however.
"While we continue the ongoing investigation into the serious allegations concerning Kevin Spacey’s behavior on the set of 'House of Cards,' he has been suspended, effective immediately," Media Rights Capital, the company that produces the show, said in a statement.
"MRC, in partnership with Netflix, will continue to evaluate a creative path forward for the program during the hiatus," it said.
The Netflix statement also said the company will work with MRC to determine the show's future.
Spacey, who has had a successful career in film, television and theater, is one of several big names in the entertainment business to be accused of sexual misconduct by people coming forward in the wake of allegations against movie producer Harvey Weinstein.
Spacey apologized last weekend to actor Anthony Rapp, who had accused him of trying to seduce him in 1986 when Rapp was 14. CNN reported on Thursday that eight current and former employees of "House of Cards," who were not identified, alleged sexual misconduct against Spacey.
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