- Shameik Moore
- Miles Morales
- Hailee Steinfeld
- Gwen Stacy / Spider-Woman (voice)
- Brian Tyree Henry
- Jefferson Davis
- Nicolas Cage
- Spider-Noir
- Luna Lauren Velez
- Rio Morales
- Jake Johnson
- Peter B. Parker
- Oscar Isaac
- Miguel O'Hara
- Jason Schwartzman
- Jonathan Ohn / The Spot (voice)
- Issa Rae
- Jessica Drew / Spider-Woman (voice)
- Karan Soni
- Pavitr Prabhakar / Spider-Man India (voice)
- Amandla Stenberg
- Margo Kess / Spider-Byte (voice)
- John Mulaney
- Spider-Ham
- Kimiko Glenn
- Peni Parker
- Daniel Kaluuya
- Hobie Brown / Spider-Punk (voice)
- Mahershala Ali
- Aaron Davis
- Jharrel Jerome
- Miles G. Morales / The Prowler (voice)
