May Parker (aunt, née Reilly) — Peter Parker's loving aunt, who raises him after his parents died. After the murder of her husband, Ben (Peter's Uncle), May is virtually his only family, and they are very close.
Zoe Parker (Cousin) - The history of the character is unknown
Mary Jane Watson-Parker (former wife) — originally merely Gwen Stacy's competition, M.J. eventually became Peter's best friend and wife. After Peter and Mary Jane made a deal with Mephisto to save Aunt May's life, their marriage is erased from official continuity.
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)
May Parker's Family
Albert Reilly (father)
Claire Reilly (mother)
Horace Reilly (uncle)
Bill Reilly (uncle)
Claudia Reilly (aunt) — Bill Reilly's wife
Sam Reilly (cousin) — Bill and Claudia's son
Julia Reilly (cousin) — Bill and Claudia's daughter (married last name unrevealed)
Jan Reilly (sister)
April Reilly (sister)
Alexa (niece) — Julia Reilly's daughter and Peter Parker's cousin, last name unrevealed
Ames (niece) — Peter Parker's cousin, last name unrevealed
Amanda (niece) — Peter Parker's cousin, last name unrevealed
Peter Parker's love interests
Gwen Stacy — Gwen was Peter's first serious girlfriend. She was very kind but slightly spoiled, smart, beautiful and shared Peter's love for science. Her father was police Captain George Stacy. Peter initially ignored her, due to his concern for his sick Aunt May, which frustrated Gwen. First a friendship, then a romance gradually formed between the two, which lasted for over a year, until her death. She was killed by the Green Goblin when he threw her off a bridge. In House of M, Gwen is still alive and married to Peter.
Betty Brant — Betty takes over her mother's former position as Daily Bugle secretary after she dies. Peter dates Betty Brant for a while, but they break up due to her blaming Spider-Man for the death of her brother. She later marries Daily Bugle reporter Ned Leeds, although she briefly gets back with Peter after the breakdown of her marriage. Despite this, both Betty and Peter maintain a close friendship.
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. She had also been friends with Gwen Stacy. At Harry Osborn's goodbye party Peter asks her to be his girlfriend and the two share their first kiss. However they break up after Spider Island due to surmising that Peter was Spider-Man, and was angry that he'd lied to her.
Mary Jane Watson: listed above. A former shallow, sexy party-girl who matured and settled down with Peter eventually after Gwen (with whom Mary Jane was friends) died.
Black Cat a.k.a. Felicia Hardy — Peter's first superhero girlfriend. She is calculating, strong and sly. Felicia competed against Mary Jane for Peter's affections, eventually losing after she left town for a while.
Michele Gonzales — Michele is a criminal defense lawyer and the hot-tempered sister of Peter's roommate Vin Gonzales, as well his temporary roommate while Vin serves time for his involvement in the Spider-Tracer Killings frame-up (to which she got him a plea bargain). When Michelle attempts to kick Peter out (who is actually the Chameleon in disguise) she is instead seduced by him and become infatuated with him. She is almost constantly angry, and questioning her or drawing attention to the size of her buttocks really sets her off. However,she is also a helpful and kind person, by trying hard to help her clients get their lives back on track. She returned to her previous home in Chicago shortly after Vin's release.
Ms. Marvel — Carol Danvers, the superhero Ms. Marvel, has worked with Spider-Man on occasion and even agreed to go on a date with him in accordance with his helping her on a mission, despite how angry he can make her. She later fulfills her promise. Spider-Man had admitted to himself he finds her attractive in her outfit. At the end of the near disastrous date, the two bonded together over a love of junk food. After she was possessed by the symbiote for a time, Venom hints to Spider-Man that his feelings for Ms. Marvel are mutual. The two have remained close friends.
Liz Allan — Peter's high school crush, although they never get together. Liz instead marries Peter's friend, Harry Osborn.
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. 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 strong feelings for her.
Kitty Pryde — In the Ultimate Marvel Universe, Kitty Pryde of the X-Men had a crush on Spider-Man and would eventually become his girlfriend.
Sophia "Chat" Sanduval — A mutant known as Chat from the Marvel Adventures Spider-Man who appeared in issue 53 and is best friends with this version's Emma Frost. She has the power to talk to animals. She is one of the few people who knows Spider-Man's identity and has developed deep feelings for him. She and Peter begin dating, meanwhile Emma (with her own crush on Peter) shakes their relationship. Eventually, Chat becomes Peter's girlfriend.
Silver Sable — In the What If? storyline What If Spider-man had married the Black Cat?, after Black Cat is accidentally killed by the Paladin, Silver Sable and Spider-Man enter into a romantic relationship due to their shared grief over the loss of their loved ones; Uatu the Watcher leaves it ambiguous whether this relationship will work out or not. Incidentally, the 616 version of Sable recently displayed romantic interest towards Spider-man.
Emma Frost — In Marvel Adventures Spider-man, Emma reveals that she has a crush on Peter and tries to separate Chat and Spidey by erasing Chat's memory. On her own, she decides to pay for her crimes and get arrested, but returns to help Spidey, apparently, still holding a crush on Peter.
Rogue — In X-Men Forever continuity, Spider-Man helped Rogue when she was annoyed with events happening around herself, her new powers and the X-Men's current status. Rogue somewhat takes Spider-Man's advice to follow her heart and, now that she can touch people, almost kisses him.
Ben Reilly's love interests
Janine Godbe aka Elizabeth Tyne The one great love of Ben's life. Ben found out Janine was living under an assumed name after she murdered the father who molested her. Ben confided in Janine that he was a clone of Spider-Man. Later Kaine forced Janine to fake her death to hurt Ben. Janine later turned herself into the police for killing her father. In the MC2 universe Darkdevil is Ben and Janine's son.
Betty Brant was attracted to the Scarlet Spider during the Web of Life storyline where Ben saved her from the Grim Hunter.
Jessica Carradine is the daughter of the burglar who killed Peter Parker's uncle Ben. She dated Ben during the period where he was Spider-Man and thought he was the real Peter Parker. Janine is a photographer and discovers Ben is Spider-Man.
Firestar — Ben and Firestar both had an attraction towards each other, but it was were never acted upon.
Lois Lane — When Ben competed in Marvel vs DC he met Lois Lane and asked her on a date to find out she is engaged to Clark Kent/Superman
Carrie Bradley a woman Ben was supposed to go on a date with but failed to make the date on time because of his life as a super hero.
Black Cat — They both dated when Ben was acting as Spider-Man. He admitted that he was not the Spider-Man she knew, but he never revealed his true identity.
Ben had Peter's memories of dating Betty Brant and Gwen Stacy, though not Mary Jane. This is due to the fact that Peter had not yet developed feelings for Mary Jane, at the time he was cloned.
Eddie Brock — An enemy of Spider-Man's, who formerly wore the Venom and Anti-Venom symbiotes, but teams up with him on numerous occasions to fight Carnage.
Jean DeWolff — A police captain and good friend of Spider-Man. She fell in love with Spider-Man (rather than Peter Parker, as she never learned his true identity), and eventually confessed those feelings to him during a time when he was physically asleep and his body being manipulated by the Venom symbiote. She eventually died in the line of duty, with Peter never knowing that she had had feelings for him, though he did have suspicions after finding a collection of photographs of the two of them together after her death.
Glory Grant — Former neighbor and friend of Peter Parker. Later becomes J. Jonah Jameson's secretary.
Gabrielle Greer — A young woman who lived at the same hotel as Ben, when he first returned to New York City.
Shirley Lewis (née Washington) — Owner of the Daily Grind and Ben's boss.
Desiree Winthrop — A model and regular at the Daily Grind.
Buzz — A regular at the Daily Grind (his real name was never stated). On a few occasions he helped Ben (not realizing he's Spider-Man), through useful advice.
Devon Lewis — Son of Shirley and part-time employee at the Daily Grind. At first Devon was distrustful of Ben, but over the months that followed he warmed up to him.
Kaine's supporting characters
Aracely — A 16 year old Mexican girl, whom Kaine saved from human traffickers and took in, upon his arrival in Houston. It's later revealed that Aracely possesses psychic abilities, similar to Madame Web. Her full name is María Aracely Josefina Penalba de las Heras, and adopted the codename "Hummingbird"
Annabelle Adams — A bartender/singer at the Four Seasons Kaine lives at. Annabelle is Mexican, which prompted Kaine to ask her for help with Aracely (who couldn't speak English). Kaine later attends a concert Annabelle's band holds.
Donald Meland — A Houston doctor whom Kaine meets when he brings in the half-dead Aracely to the hospital.
Wallence "Wally" Layton — A Houston police officer, who encouraged Kaine to continue as a vigilante in Houston. Wally is gay and married to Donald Meland.
Gwen Stacy's family
George Stacy (deceased) — Gwen Stacy's father, Police Captain. Introduced in The Amazing Spider-Man (vol. 1) #56 (1968). He approves of Peter and Gwen's relationship as boyfriend and girlfriend. During a fight between Spider-Man and Doctor Octopus, he is crushed by falling debris while saving a child. As he dies, he reveals to Peter that he had known his identity for some time (something Peter had suspected anyway), and asks Peter to take care of Gwen.
Helen Stacy — Gwen's mother and wife of George. Her first and only appearance is Peter Parker: Spider-Man #-1 (minus one).
Arthur Stacy — Gwen Stacy's uncle, a private investigator, first appeared in The Amazing Spider-Man (vol. 1) #93 and #95. He was reintroduced only in the 1990s, in Peter Parker: Spider-Man #70 (in the last part of 'Clone Saga'). He is George's younger brother, but was originally presented in the 1970s as the older brother. For a time, Spider-Man would call on Stacy's skills as an investigator.
Nancy Stacy — Gwen's aunt. Wife of Arthur and mother of Jill and Paul. First appeared in The Amazing Spider-Man (vol. 1) #93.
Jill Stacy — Arthur Stacy's daughter and Gwen's cousin. Friend of Mary Jane and Peter. After Mary Jane was kidnapped, and thought to be dead, Jill made romantic advances toward Peter. Introduced in Peter Parker: Spider-Man #76 (1997).
Paul Stacy — Arthur Stacy's son and Gwen's cousin. He blames Spider-Man for Gwen's death and joins the anti-mutant group the Friends of Humanity in an attempt to bring him to justice. Introduced in The Amazing Spider-Man (vol. 1) #422.
Gabriel (last name unrevealed) — Gwen's son by Norman Osborn. Norman convinced Gabriel and his sister, Sarah, that Peter Parker was their father and had killed their mother. Although Sarah is persuaded otherwise, Gabriel continues to believe so and takes on the identity of the "Gray" Goblin. After a confrontation with Spider-Man, he crashed into the river on his glider and lost his memories. Sarah took him to their home in France to recover. After failing to convince Sarah to join him, he flies off on a different glider. Introduced in The Amazing Spider-Man #509.
Sarah (last name unrevealed) — Gwen's daughter by Norman Osborn. Norman convinced Sarah and her brother, Gabriel, that Peter Parker was their father and had killed their mother. Sarah becomes suspicious after she meets Spider-Man however. She is convinced of the truth when Spider-Man saves her life by giving her a blood transfusion after she is shot by police. Spider-Man later learns that the pain caused by her accelerated aging has led her to abusepainkillers, and her addiction has gotten her in trouble with the French authorities. However, she promises to seek help, and perhaps someday become a hero herself. Introduced in The Amazing Spider-Man #509.
Anthony "Henry" Harper (appeared in The Amazing Spider-Man, voiced by David Lodge) — Nothing much is known about District Attorney Henry Harper's past. In one side-mission in the game, he is kidnapped by Iguana and is dragged into the sewers through the train docking station. After saving a civilian, Spider-Man learns that Harper was a pawn for Oscorp. After Spider-Man defeats Iguana, Harper is rescued and he escapes out of the sewers. Sometime before the events of the game, Harper had focused on exposing Quest Aerospace's evil schemes after he successfully prosecuted some of the city's most notorious criminals. The corrupt corporation lost millions of dollars to Harper, but they fired back when they had evidence of funds contributed to the D.A.'s reelection campaign were sourced by Oscorp Industries. These allegations were never revealed, but Harper's reputation was severely damaged in the eyes of many citizens.
Dr. Watts (appeared in Spider-Man 2: Enter Electro) — Dr. Watts (first name unknown) is a prominent scientist. She is the creator of the Bio-Nexus Device and a world-renowned scholar of biology. In the game, she is kidnapped by Hammerhead while attending the Science Industry Ball. During Spider-Man's fight with Hammerhead, Dr. Watts disappears, which led Spider-Man to believe that the Sandman took her. Due to an anonymous tip, Spider-Man tracks Dr. Watts to a museum, where she is taken hostage by Electro. After Electro leaves the museum, Dr. Watts tells Spider-Man that Electro went to the Twin Towers.
Aaron Warren (appeared in The Spectacular Spider-Man) — Mr. Warren is Midtown High's biology teacher. He considers Peter and Gwen his smartest students, and recommended them to be interns at Empire State University.
Detective Terri Lee (appeared in Spider-Man) — She is a detective for the New York Police Department. Naturally, she investigates cases that involve Spider-Man and to an extent to Peter Parker, though she never found out Peter and Spider-Man are the same person. She didn't trust Spider-Man at first, but overtime she began accepting Spider-Man as an ally. She has ties to Carnage and she had a relationship with the vampire hunter Blade.
Indira "Indy" Daimonji (appeared in Spider-Man: The New Animated Series) — She is Peter Parker's potential love interest and a rival of Mary Jane Watson. She works at a news network down in Manhattan. She is seriously injured by Spider-Man in the series finale when Spider-Man is tricked by the Gaines Twins into injuring Indy. This led to Peter giving up being Spider-Man
Whitney Chang (appeared in The Amazing Spider-Man, voiced by Claudia Black) — She is a top investigative reporter for the Channel 3 News Network and is well known for putting herself at risk for finding out the truth. She met Spider-Man in person while she was investigating a secret Oscorp facility involving cross-species genetics and its connection with Alistair Smythe, with Spider-Man tracking down a crate with Dr. Connor's research to create the cure. Whitney gave Spider-Man her camera and asks him to take photos exposing Oscorp's research. As explained in her bio, Chang grew up in the Sunset Park region of Brooklyn, New York, where she witnessed a neighbor being murdered by an angry mob after being framed for murder and being slandered by the news for days. After graduating from Yale, she quits her job as the host of a music video channel and snuck aboard a flight to Iraq, arriving just as American troops invade Baghdad.
Enemies in other media
Monster-Ock (appeared in Spider-Man, with vocal effects provided by Marcus Shirock) — He is an exclusive villain to the game. When Spider-Man defeats Carnage by using sonic weaponry, the symbiote left Cletus Kasady and it bonds onto an unconscious Doctor Octopus and Monster-Ock attempts to kill Spider-Man. After a lengthly chase throughout the underwater base, Spider-Man barely escapes and the symbiote left Octavius, leaving him catatonic, but alive. Spider-Man saves Octavius and the symbiote's, as well as Kasady's, fates are unrevealed.
Talon (appeared in Spider-Man: The New Animated Series, voiced by Eve) — She is a villain created exclusively for the show. She bears many characteristics to the Black Cat. In her first and only appearance, "Keeping Secrets", she is behind a string of robberies, much to Spider-Man's dismay, who is a little concerned for her safety. Her real name is Cherry Valentine, and she is Harry Osborn's girlfriend. Peter learns about this, but he keeps this a secret to both Mary Jane and Harry. During a later encounter, she is shot by the police and she is seemingly killed. At the end of the episode, it was revealed she was alive and well, and she breaks up with Harry.
Black Cat 2099 (appeared in Spider-Man: Edge of Time, voiced by Katee Sackhoff) — She is a clone of the original Black Cat. It was revealed that she was upgraded with cybernetic weaponry and she claimed that Alchemax was forcing her to use anti-aging drugs to mislead Spider-Man. Black Cat 2099 also has clones of herself.
Walker Sloan (appeared in Spider-Man: Edge of Time, voiced by Val Kilmer) — Walker Sloan is one of Alchemax's greatest minds, and he serves as one of the main villains in the game. In the game, he travels back in time to establish Alchemax sometime in the 1970s, which results in Spider-Man's untimely death by the hands of Anti-Venom, who is revealed to be under mind control. Sloan was revealed to be working with Doctor Octopus to alter reality, much to Octavius' dismay. After Anti-Venom was freed from mind control, he prepares to kill Sloan for mindwiping him. Instead, Anti-Venom, Sloan, and Octavius were pulled into an unstable portal, resulting in the creation of Atrocity.
Atrocity (appeared in Spider-Man: Edge of Time, with vocal effects provided by Fred Tatasciore) — Atrocity is a result of the violent fusion of Anti-Venom, Doctor Octopus, and Walker Sloan. Spider-Man and Spider-Man 2099 are unaware of its identity, until Spider-Man collects three samples of its DNA and analyzed it to Spider-Man 2099. In the end, Spider-Man defeats Atrocity by luring him into a portal, and Anti-Venom, Sloan, and Octavius' fates are unknown.