The seven samurai
- Takashi Shimura as Kambei Shimada (島田勘兵衛, Shimada Kanbei), a war-weary but honourable and strategic rōnin, and the leader of the seven.
- Yoshio Inaba as Gorōbei Katayama (片山五郎兵衛, Katayama Gorōbei), a skilled archer, who acts as Kambei's second-in-command and helps create the master plan for the village's defense.
- Daisuke Katō as Shichirōji (七郎次), Kambei's old friend and former lieutenant.
- Seiji Miyaguchi as Kyūzō (久蔵), a serious, stone-faced and supremely skilled swordsman.
- Minoru Chiaki as Heihachi Hayashida (林田平八, Hayashida Heihachi), an amiable though less-skilled fighter, whose charm and wit maintain his comrades' morale in the face of adversity.
- Isao Kimura as Katsushirō Okamoto (岡本勝四郎, Okamoto Katsushirō), the untested son of a wealthy landowner samurai, who Kambei reluctanctly takes in as a disciple.
- Toshiro Mifune as Kikuchiyo (菊千代), a humorous, mercurial and temperamental rogue who lies about being a samurai, but eventually proves his worth and resourcefulness.
- Yoshio Tsuchiya as Rikichi (利吉), a hotheaded and relatively young villager
- Bokuzen Hidari as Yohei (与平), a very timid old man who shares some comic scenes with Kikuchiyo
- Yukiko Shimazaki as Rikichi's wife
- Kamatari Fujiwara as Manzō (万造), a farmer who disguises his daughter as a man to try to protect her from the samurai
- Keiko Tsushima as Shino (志乃), Manzō's daughter
- Kokuten Kōdō as Gisaku (儀作), the elder miller and village patriarch, referred to as "Grandad"
- Yoshio Kosugi as Mosuke, one of the farmers sent to town to hire samurai
- Shinpei Takagi as the bandit chief
- Shin Otomo as the bandit second-in-command
- Haruo Nakajima as a bandit
- Eijirō Tōno as a thief
- Atsushi Watanabe as a bun seller
- Jun Tatara as a coolie
- Sachio Sakai as a coolie
- Takeshi Seki as a coolie
- Tatsuya Nakadai as a samurai wandering through town (uncredited)
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