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• Sigourney Weaver as Ellen Ripley (officially designated as Ripley 8), reprising her role from the previous three Alien films. After having sacrificed herself to kill the Alien queen gestating inside her in Alien 3, Ripley has been cloned using blood samples so that the military may extract the queen embryo. As a result of the cloning process Ripley has been affected by the Alien queen's DNA. She has enhanced strength and reflexes, acidic blood, and the ability to sense the presence of the Aliens. Weaver also portrayed the failed seventh clone.

• Nicole Fellows as Young Ripley

• Winona Ryder as Annalee Call, the newest crew member of the Betty. She recognizes Ripley and has knowledge of the Aliens. Call is revealed during the course of the film to be a synthetic and helps the surviving crew interface with the Auriga.

• Dominique Pinon as Dom Vriess, the Betty's mechanic. A paraplegic, he uses a motorized wheelchair. Vriess shares a close friendship with Call and an antagonistic relationship with Johner.

• Ron Perlman as Ron Johner, a mercenary and member of the Betty's crew. Johner makes bad jokes, has a short temper, and teases Vriess about his handicap.

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