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More than a few characters in this game have become audience all-stars.
Familiars
A good number of the game's familiars have received their own popularity, in some even cases moreso than playable characters.
- Dark Bum
. Introduced in Rebirth, he quickly became one of the most wanted items, not just because of how cute he looks, but also because of his borderline Game-Breaker power, turning red hearts into soul hearts. Although the first nerf was somewhat minor, when he was severely nerfed to have a chance to spawn spiders, there was fan outcry for him. Nevertheless, Dark Bum still remains one of the most popular and loved familiars overall. - Sissy Longlegs, another familiar from Rebirth. While not a powerful item, like Dark Bum, Sissy is considered cute in her own way. Her item description "She loves you" and respectable ability tend to give her a free pass on rerolls. Chanting her name when one gets her isn't unheard of, either.
- Bumbo from Afterbirth has won a lot of fans over with his array of rather useful abilities. The fact that he grows by collecting money, which tends to be rather useless later on, also helps. Edmund seems to have been very aware of his popularity, as he even received his own Spin-Off game.
- Dry Baby has a good number of fans, due to its ability to block pretty much all shots and damaging every enemy in the room each few shots it blocks.
- Psy Fly, due to being a Game-Breaker that turns every bullet-hell fight into a complete joke. It got to the point where when it received a minor nerf in the November 2021 update, the patch notes went out of their way to reassure fans it was still really good.
Bosses
- The Seven Deadly Sins tend to get a lot of fanart, including Greed (in spite of his Scrappy status when it comes to the actual game).
- Among the devil-like boss monsters, Dark One and Adversary seem to be the most popular.
- As for the more "normal" bosses (if one can call them normal), Monstro, Chub, and Loki have always remained fan-avorites.
- The Rainmaker, the flamenco-dancing, water-bending poltergeist boss, earned quite a few fans for his quirkiness. Being adorable also helps. He's so likable, Edmund said he wanted to cut him from Repentance for not fitting the Isaac universe but couldn't bring himself to do it.
- The Siren earned a fair amount of popularity, partly for her unique gimmick of stealing your familiars and partly due to being deemed a Cute Monster Girl by players.
- The Visage is another well-liked battle for being a very well-executed Dual Boss. The two parts of The Visage will fight you in tandem and interact with each other in a lot of their attacks, and unlike Mask of Infamy, you have to constantly deal with both of them instead of simply defeating them one at a time and the Heart actually puts up a good fight with a number of tricky attacks across three phases.
- The Scourge is gaining a following for its Ugly Cute design and tricky fight.
- Rotgut. It starts out normal enough, with you fighting a Sarlacc-esque monster that has admittedly tricky-to-dodge attacks, but once you deplete its health, it sucks you into it and the fight continues inside of it, in a 2D Crawl-space battle! First, you have to kill a giant maggot inside of it, and then you have to kill its testicle-like organs; killing both causes it to spit you back out, and ends the fight.
- Maid in the Mist is shockingly popular despite not even existing in the game! She's the beta design for Min-Min, and all that remains of her is a boss portrait in the game files, but that was enough for fans to fall in love with her.
