 | Hudson Thames | | Peter Parker / Spider-Man |
| Eugene Byrd | | Lonnie Lincoln |
| Kari Wahlgren | | May Parker / Roxanna Volkov / Mrs. Eberhardt |
| Colman Domingo | | Norman Osborn |
| Cathy Ang | | Pearl Pangan |
| Grace Song | | Nico Minoru |
| Erica Luttrell | | Asha / Mrs. Lincoln / Emma |
| Zeno Robinson | | Harry Osborn / Mr. Campbell |
| Leilani Barrett | | Big Donovan |
| Roger Craig Smith | | Coach Grayfield / Dmitri Smerdyakov / Chameleon / James Sanders |
| Ettore Ewen | | Bulldozer |
| Zehra Fazal | | Dr. Connors / Susan O'Hara / Hot Rod |
| Hugh Dancy | | Dr. Otto Octavius |
| Anjali Kunapaneni | | Jeanne Foucault / Finesse |
| Anairis Quinones | | Carmilla Black / Maria Vasquez / Ms. Anand |
| Paul F. Tompkins | | Bentley Wittman |
| Aleks Le | | Amadeus Cho |
| Jonathan Medina | | Mac Gargan |
| Travis Willingham | | Mikhail Sytsevich / Thaddeus Ross |
| Phil LaMarr | | Mr. Lincoln / Mr. Taylor |
| Kellen Goff | | Symbiotic Alien / Vincent Patillo |
| Alex Désert | | Grift |
| Charlie Cox | | Daredevil |
| Matte Martinez | | Andre Lincoln |
| Robin Atkin Downes | | Doctor Strange / Dr. Strange |
| Sarah Natochenny | | Mila Masaryk |
| Ogie Banks | | Pizza Shop Owner |
| Zach Cherry | | Klev |
| Jake Green | | Butane |
| Mick Wingert | | Iron Man / Tony Stark |
