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Note: For this list only the creators of the characters first appearances are listed. As with all comic book characters Spider-Man and his supporting cast have had several reinventions and different contributions from different writers. Theses include different iterations in different mediums that all have added different concepts to the overall mythology of the characters.

Theirs Peter Benjamin Parker/Spider-Man, referred to as Peter Parker or Spider-Man on screen. He made his first appearance in the comic story "Spider-Man!" from Amazing Fantasy #15 (June 1962) by editor/writer Stan Lee and writer/artist Steve Ditko.

Norman Virgil Osborn/The Green Goblin, referred to as Norman Osborn or Green Goblin on screen. He made his first appearance as Green Goblin in the comic story "The Grotesque Adventure of the Green Goblin" from The Amazing Spider-Man #14 (July 1964) and he also made his first appearance as Norman Osborn in the comic story "The Goblin and the Gangsters" from Amazing Spider-Man #23 (April 1965). The character was created by editor/writer Stan Lee and writer/artist Steve Ditko.

Mary Jane "MJ" Watson, who made her first appearance in the comic story "Captured By J. Jonah Jameson!" in Amazing Spider-Man #25 (June 1965) in a cameo appearance followed by a full appearance in the comic story "The Birth of a Super-Hero!" from Amazing Spider-Man #42. The character was created by editor/writer Stan Lee and artist John Romita Sr..

Harold Theopolis "Harry" Osborn, referred to as Harry Osborn on screen. He made his first appearance in the comic story "If This Be My Destiny...!" from The Amazing Spider-Man #31 (December 1965) by editor/writer Stan Lee and writer/artist Steve Ditko.

Peter's Uncle Benjamin Franklin "Ben" Parker, referred to as Uncle Ben or Ben Parker on screen. He made his first appearance in the comic story "Spider-Man!" from Amazing Fantasy #15 (June 1962) by editor/writer Stan Lee and writer/artist Steve Ditko.

Peter's Aunt Maybelle "May" Parker, referred to as Aunt May or May Parker on screen. She made his first appearance in the comic story "Spider-Man!" from Amazing Fantasy #15 (June 1962) by editor/writer Stan Lee and writer/artist Steve Ditko.

John Jonah Jameson, Jr., referred to as J Jonah Jameson on screen. He made his first appearance in the comic story "Spider-Man" from Amazing Spider-Man #1 (March 1963) by editor/writer Stan Lee and writer/artist Steve Ditko.

Eugene "Flash" Thompson, referred to as Flash Thompson on Screen. He made his first appearance in the comic story "Spider-Man!" from Amazing Fantasy #15 (June 1962) by editor/writer Stan Lee and writer/artist Steve Ditko.

Joseph "Robbie" Robertson, referred to as Robbie or Robbie Robertson on screen. He made his first appearance in the comic story "In the Clutches of the Kingpin!" from Amazing Spider-Man #51 (August 1967) by editor/writer Stan Lee and artist John Romita Sr..

Dr. Mendel Stromm, who made his first appearance in the comic story "Once Upon A Time, There Was A Robot...!" from The Amazing Spider-Man #37 (June 1966) by writer Stan Lee and artist Steve Ditko.

The Carjacker who kills Uncle Ben, called The Burgler in the comics. He made his first appearance in the comic story "Spider-Man!" from Amazing Fantasy #15 (June 1962) by editor/writer Stan Lee and writer/artist Steve Ditko.

Bonesaw McGraw, called Joseph "Crusher" Hogan in the comics. He made his first appearance in the comic story "Spider-Man!" from Amazing Fantasy #15 (June 1962) by editor/writer Stan Lee and writer/artist Steve Ditko.

and Elizabeth "Betty" Brant, referred to Betty Brant on screen. She made her first appearance in the comic story "Nothing Can Stop... the Sandman!" from The Amazing Spider-Man #4 (September 1963) by writer Stan Lee and writer/artist Steve Ditko.

All other characters were created for the movie by the films writers.

No. Brock, who becomes the supervillain Venom in the comics (and the 2007 sequel) is referenced in dialogue ("Eddie's been on it for weeks..."), but does not appear. The character had scenes filmed, but those scenes were cut from the final film, and the deleted scenes do not appear on the DVD. Brock appears later in Spider-Man 3 (2007).

Bluto hangs from a tree and kisses Olive Oyl the same way in the Popeye short, Puppet Love (1944).

Spider-Man kisses Felicia Hardy upside down, in the balcony of her house in Spider-Man (1994).

Yes, though the film took several elements from the Ultimate Spider-Man series, which is an updated retelling of the original stories from the sixties. As such, this list points out the differences between the original stories: (1) In the comics, Peter is bitten by a radioactive spider that was caught in a beam of energy during a science experiment, rather then the genetically altered "super spider" in the film, (2) Peter did not know Harry Osborn or Mary Jane Watson in high school; he met them while at college, along with Gwen Stacy, (3) Peter's first love interest was Betty Brant of the Daily Bugle and then Gwen Stacey (Mary Jane didn't come into the picture until later); (4) Peter's spider-sense does not allow him to precisely see threats to him; it just creates a tingling sensation that varies in intensity depending on the severity of the danger detected, (5) Uncle Ben is killed by a burglar, as opposed to the carjacker in the film, (6) After Peter won the wrestling match, he went on to become a TV sensation, appearing on various talk shows and programs; in the film, he is stiffed of his money by the promoter, and does not go on to TV, and (7) Spider-Man uses lots of wisecracks and sarcasm during encounters with his enemies; in this film and in the two sequels, Spidey's sense of humor is only occasionally represented. The biggest change of all is undoubtedly the organic web shooters, as opposed simply to the wrist mounted guns Peter created that used a special formula he created.