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Nagakura Shinpachi:
Complete Chapter 5 to unlock him in two player mode.

Okita Souji:
Complete Okita Souji's story mode quest to unlock him in two player mode.

Saigo Takamori:
Complete Saigo Takamori's story mode quest to unlock him in two player mode.

Saito Hajime:
Complete Saito Hajime repeatedly on Te-awase to unlock him in two player mode.

Sakamato Ryoma:
Complete Sakamato Ryoma's story mode quest to unlock him in two player mode.

Serizawa Kamo:
Complete Serizawa Kamo's side quest to unlock him in two player mode.

Shinsengumi Lower Class member:
Start a game on the Shinsengumi storyline to unlock this character in two player mode.

Takeda Kanryuusai:
Complete the mission with Takeda Kanryuusai at Masuya to unlock him in two player mode.

Tetsunosuke Ichimura:
Meet Tetsunosuke Ichimura during Chapter 6 of the Shinsengumi storyline to unlock in two player mode.

Toudou Heisuke:
Kill Toudou Heisuke at the end of Chapter 4 to unlock him in two player mode.

Two Hitokiri:
Complete the story mode for Two Hitokiri to unlock him in two player mode.

Yamanami Keisuke:
Kill Yamanami Keisuke at the end of Chapter 3 to unlock him in two player mode.

Yamazaki Susumu:
Start a game in the Shinsengumi storyline to unlock him in two player mode.

School power-up items:

Choshu Ryu: In the Imperial storyline, get all techniques of the school then defeat the sword master.

Edo Ryu: In the Imperial storyline, get all techniques of the school then defeat the sword master.

Hokushin Ittou Ryu: In either storyline, get all techniques of the school then defeat the sword master.

Satsuma Ryu (Jigen Ryu): In the Imperial storyline, get all techniques of the school then defeat the sword master.

Shintou Munen Ryu: In either storyline, get all techniques of the school then defeat the sword master.

Spear Ryu (Houzouin Ryu): In the Shinsengumi storyline, get all techniques of the school then defeat the spear master.

Tama Ryu (Tennen Rishin Ryu): In the Shinsengumi storyline, get all techniques of the school then defeat the sword master before Chapter 6.

Hint: Better chance of winning:
Ride a horse that is "A" or "B" rated with a lot of stamina that prefers to be at the back of the pack. When about 5 furlongs away from the finish line, press Triangle to get closer to the front. Use most or all of your stamina before letting the horse run out to the leaders.

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