The leader of Yorozuya, who will do anything for a price. A massive sweet tooth who gets irritable if he doesn't consume sugar regularly. Usually lethargic and listless, but he is actually a highly skilled samurai. A former Joui patriot once feared as the "White Yaksha" for his swordsmanship, he now uses only a wooden sword. He has silver, naturally permed hair. He speaks in an aloof, detached manner with a slight Edo-style roughness. First person: I (rough, casual).

The son of a swordsmanship dojo owner whose legacy was ruined by the sword ban following his father's death. After Gintoki saved his sister from debt collectors, he began working at Yorozuya to learn Gintoki's spirit. However, he soon came to regret that decision... He is the primary straight man of the series, though he has been known to play the fool lately.

A girl from the Yato Tribe, the strongest combat race in the universe. She became a self-invited part-timer at Yorozuya to save money to return to her home planet. Despite her cute appearance, she is a glutton, violent, and sharp-tongued.

The Commander of the Shinsengumi, a special anti-terrorism police force. He is the man who organized the Shinsengumi, and he enjoys deep trust from his subordinates. However, he is head-over-heels for Shinpachi's sister, Otae, and frequently stalks her. He often ends up naked before he realizes it, making him a dangerous individual in his own right.

The cool-headed Vice-Commander of the Shinsengumi. He established the Kyokuchu Hatto (Code of Conduct), and once he draws his blade, he is feared by terrorists and his own men alike as the "Demon Vice-Commander." He is a heavy smoker and a severe mayonnaise addict.

The Shinsengumi's best swordsman. Contrary to his refreshing appearance, he possesses a dark, sadistic interior. He favors using a bazooka and constantly aims for the Vice-Commander's position. He looks for any excuse to slack off on duty. He is a total sadist. He speaks in a detached, aloof manner with a distinct Edo dialect. Second person: you (blunt).

A dangerous man who views the world ruled by the Amanto as his enemy and plots terrorism leading the armed group Kiheitai. He possesses a mysterious charisma that draws people to him. Honestly, he has no interest in Matako.

A radical Joui extremist who fought alongside Gintoki and the others during the Joui War (the battle against the Amanto). He continues to work toward driving out the Amanto and rebuilding the country of samurai. His nickname is Zura.

A comrade who fought with Gintoki and the others during the Joui War. He possesses the spirit to pursue a grand cause without sweating the small stuff, and he has gathered many followers.

The lone female member of the Kiheitai, who is in love with Takasugi. She is a master of dual pistols and is known by the alias "Red Bullet." She tries to hug Takasugi whenever she sees an opening.