Neil Patrick Harris
Peter Parker / Spider-Man (voice) (13 eps)
Lisa Loeb
Mary Jane Watson (voice) (13 eps)
Ian Ziering
Harry Osborn (voice) (13 eps)
Keith Carradine
J. Jonah Jameson (voice) (5 eps)
Ed Asner
Officer Barr (voice) (4 eps)
Rino Romano
Muang / Drip Den guy (voice) (4 eps)
Ethan Embry
Max Dillon / Electro (voice) (3 eps)
Julie Nathanson
Sally Johnson (voice) (3 eps)
Virginia Madsen
Silver Sable (voice) (2 eps)
Michael Dorn
Kraven the Hunter (voice) (2 eps)
James Marsters
Sergei (voice) (2 eps)
Clancy Brown
Raymond (voice) (2 eps)
Jeffrey Combs
Dr. Zellner (voice) (2 eps)
Karen Maruyama
Head Protestor (voice) (2 eps)
Jeremy Piven
Roland Gaines (voice) (2 eps)
Keith David
Agent Mosely (voice) (2 eps)
Kathy Griffin
Roxanne Gaines (voice) (2 eps)
Jennifer Hale
Female News Announcer (voice) (2 eps)
Angelle Brooks
Indira 'Indy' Daimonji (voice) (1 eps)
Michael Clarke Duncan
Wilson Fisk / The Kingpin (voice) (1 eps)
Stan Lee
Frank Elson (voice) (1 eps)
Rob Zombie
Dr. Curt Connors / The Lizard (voice) (1 eps)
Gina Gershon
Shikata (voice) (1 eps)
John C. McGinley
Richard Damien (voice) (1 eps)
Jane Lynch
Oscorp Executive (voice) (1 eps)
Tara Strong
Christina (voice) (1 eps)
Devon Sawa
Flash Thompson (voice) (1 eps)
Eve
Cheyenne Tate / The Talon (voice) (1 eps)