Friends, associates, and enemies of Spider-Man.
- Richard Parker (father, deceased)
- Mary Parker (mother, née Fitzpatrick, deceased)
- Uncle Ben Parker (uncle, deceased)
- Aunt May Parker (aunt, née Reilly)
- Mary Jane Watson-Parker (wife)
- May Parker (daughter, thought to be deceased)
- Ben Reilly aka Scarlet Spider aka Spider-Man (clone, deceased)
- Kaine (clone)
- Spidercide (clone, deceased)
- Doppelganger (mystic duplicate, deceased)
Mary Jane's family
- Anna Watson — Mary Jane's aunt and Aunt May's best friend.
- Madeline Watson (mother, deceased)
- Phillip Watson (father)
- Kristy Watson (cousin)
- Gayle Watson-Byrnes (sister)
- Timothy Byrnes (Gayle's ex-husband)
- Kevin and Tommy Byrnes (Gayle's sons)
- Frank Brown (Madeline's cousin)
- Mary MacLeod (distant aunt)
- Liz Allan — Peter's high school crush, although they never get together. Liz instead marries Peter's friend, Harry Osborn.
- Betty Brant — Peter dates the Daily Bugle secretary, but they break up due to her blaming Spider-Man for the death of her brother. Betty Brant later marries Daily Bugle reporter Ned Leeds, although she briefly gets back with Peter after the breakdown of her marriage. Despite this, both Peter and Betty maintain a close friendship.
- Gwen Stacy — Peter's first real girlfriend, whom the Green Goblin kills by throwing her off a bridge. The 2004 storyline "Sins Past" has Gwen, pregnant by Norman Osborn, secretly giving birth to twins, who, told by Norman that Spider-Man had killed their mother, attempt to kill Spider-Man in revenge as adults. In House of M, Gwen is still alive and married to Peter.
- Debra Whitman — a fellow Empire State University student and secretary whom Peter dates for a period of time, though his frequent disappearances complicate their relationship. Debra lacks self-confidence and their relationship is rather one-sided; Peter has little interest in her. She is eventually diagnosed with mild schizophrenia, ironically exacerbated by her "delusional" belief that Peter is Spider-Man. With Peter's help, she overcomes that idea. Soon afterward, she leaves New York after another man, Biff Rifkin, confesses his feelings for her.
- Marcy Kane — another Empire State University graduate student who Peter dates briefly. Prior to dating Peter, she was the lover of the Jack of Hearts. Kane is later revealed to be an alien (Contraxian) who is on Earth looking for solutions to her homeworld's problems.
- Felicia Hardy, a.k.a. Black Cat — The relationship between Spider-Man and Black Cat is short lived-after Spider-Man learns that Felicia Hardy is only interested in him as Spider-Man and not Peter Parker.
- Carlie Cooper — She is an officer of the NYPD's Crime Scene Unit and ex-best friend of Harry Osborn's ex-girlfriend, Lily Hollister. However they break up after Spider Island due to surmising that Peter was Spider-Man, and was angry that he'd lied to her. Carlie eventually left New York for her own safety.
- Sarah Rushman — Peter dates Sarah during a time when he believes Mary Jane to be dead; however, she turns out to be the S.H.I.E.L.D.-brainwashed mutant Marrow.
Ben Reilly's Love Interests
- Elisabeth Tyne, aka Janine Godbe — met Ben and lived with him for several months during his "lost years," until they were separated by Kaine. They reunited after he became Spider-Man, but were separated again when she turned herself in for her abusive father's murder.
- Jessica Carradine — photographer, daughter of the burglar who killed Uncle Ben.
- Desiree Winthrop — a wealthy young woman with limited psychic powers, of which she was unaware.
Ben also had Peter's memories of dating Liz Allan, Betty Brant, Gwen Stacy, and Mary Jane Watson.
See also: Daily Bugle
- Jean DeWolff
- Glory Grant — J. Jonah Jameson's secretary. Friend of Peter Parker.
- Crusher Hogan - the wrestler that Peter defeats shortly after gaining his powers. Later, Hogan becomes a Spider-Man fan and looks back on his encounter with the hero fondly.
- J. Jonah Jameson
- Ned Leeds
- Foggy Nelson - Matt Murdock's law partner, who briefly dated Liz Allan.
- Joe "Robbie" Robertson
- Randy Robertson — Joe Robertson's son. Friend of Peter Parker.
- Jimmy Six aka Giacomo Fortunato
- Arthur Stacy — Gwen Stacy's uncle, private investigator.
- George Stacy (deceased) — Gwen Stacy's father, Police captain, killed by falling debris in a fight between Spider-Man and Doctor Octopus.
- Jill Stacy — Arthur Stacy's daughter, friend of Mary Jane.
- Paul Stacy — Arthur Stacy's son. Blamed Spider-Man for Gwen's death and joined the anti-mutant group the Friends of Humanity in an attempt to bring him to justice.
- Flash Thompson
- Ben Urich
- MACH IV aka The Beetle
- Fantastic Four
- Gibbon (Martin Blank)
- Grizzly (Maxwell Markham)
- Human Torch (Johnny Storm)
- Steel Spider
- Madame Web
- Molten Man
- Rocket Racer
- Silver Sable
- Spider-Woman (Mattie Franklin), the niece of J. Jonah Jameson.