| Nolan North | .... | Raphael / Nightwatcher (voice) |
| James Arnold Taylor | .... | Leonardo (voice) |
| Mitchell Whitfield | .... | Donatello (voice) |
| Mikey Kelley | .... | Michelangelo (voice) |
| Mako | .... | Splinter (voice) |
| Sarah Michelle Gellar | .... | April O'Neil (voice) |
| Chris Evans | .... | Casey (voice) |
| Patrick Stewart | .... | Winters (voice) |
| Ziyi Zhang | .... | Karai (voice) |
| Laurence Fishburne | .... | Narrator (voice) |
| Kevin Smith | .... | Diner Cook (voice) |
| John DiMaggio | .... | Colonel Santino (voice) |
| Paula Mattioli | .... | General Serpiente (voice) |
| Kevin Michael Richardson | .... | General Aguila (voice) |
| Kevin Michael Richardson | .... | Additional Voices (voice) |
| Fred Tatasciore | .... | General Gato (voice) |
| Dee Baker | .... | Additional Voices (voice) |
| Greg Baldwin | .... | Additional Voices (voice) |
| Jeff Bennett | .... | Additional Voices (voice) |
| Jim Cummings | .... | Additional Voices (voice) |
| Grey Griffin | .... | Additional Voices (voice) |
| Chris Edgerly | .... | Additional Voices (voice) |
| Kim Mai Guest | .... | Additional Voices (voice) |
| Jennifer Hale | .... | Additional Voices (voice) |
| Jess Harnell | .... | Additional Voices (voice) |
| Phil LaMarr | .... | Additional Voices (voice) |
| Paul Michael Robinson | .... | Additional Voices (voice) |
| Tara Strong | .... | Additional Voices (voice) |
| Billy West | .... | Additional Voices (voice) |
| Jason Broad | .... | Background Voices (Wallah) (voice) |
| Willow Geer | .... | Background Voices (Wallah) (voice) |
| Andy Hirsch | .... | Background Voices (Wallah) (voice) |
| Barbara Harris | .... | Background Voices (Wallah) (voice) |
| Terry Maratos | .... | Background Voices (Wallah) (voice) |
| Jeremy Maxwell | .... | Background Voices (Wallah) (voice) |
| Tracy Metro | .... | Background Voices (Wallah) (voice) |
| David Michie | .... | Background Voices (Wallah) (voice) |
| David J. Randolph | .... | Background Voices (Wallah) (voice) |
| Vernon Scott | .... | Background Voices (Wallah) (voice) |
| Andreana Weiner | .... | Background Voices (Wallah) (voice) |