- Ajax threatens to sew Wade's mouth shut, which is exactly what happened to him in X-Men Origins: Wolverine.
- An action figure of the Origins version of Deadpool is seen briefly in a flashback scene.
- The line "Travel the world's exotic places, meet new and exciting people... and then kill them" was previously used verbatim by Wade Wilson in X-Men Origins: Wolverine.
- A new version of Teamheadkick's Deadpool fan rap (originally created as a tribute to the Deadpool video game) plays during the Roaring Rampage of Revenge montage.
- As Wade cuts a path through Ajax's mooks in the final battle, he stops and unmasks one he recognized: Bob, Agent of HYDRA. Rather than kill him he knocks him out and asks how the kids are doing (in that order, too).
- Worm has a blood-red eye as a reference to his cybernetic eye from the comics.
- Pre-transformation Wade Wilson is first introduced wearing a Bea Arthur T-shirt, referencing his long-time love of her in the comics.
- Near the end Deadpool tells Colossus to "go be a really big brother to someone".
- Deadpool makes a quip about the halls at Colossus's high school probably being well-monitored. This was a reference to X2: X-Men United, where Colossus's job was to guide the students down a hall to safety.
- When Colossus says he's taking Deadpool to see the Professor, Deadpool asks, "McAvoy or Stewart?", then remarks that these timelines are so confusing.
- Deadpool at one point jokes about having been bitten by a "radioactive shar-pei." This is a reference to Cable & Deadpool #2, published in 2004, where Deadpool describes himself as "Ryan Reynolds crossed with a shar-pei." Also a reference to how Spider-Man supposedly got his powers, by being bitten by a radioactive spider.
- The Bait-and-Switch scene in the beginning where Wade orders a pizza and has it delivered to a random guy's home is similar to the scene in Deadpool #10, published in 2009. The difference is that, in the comic, he kills the pizza boy for spreading a rumor about a girl so nasty that it ruined her life. Wade even orders the same toppings for the pizza, "pineapple and black olives," as well as asking if the crust was burnt.
- Negasonic mentions how often the X-Mansion gets demolished, a tendency that the actual comics were mocking by the 2000s.
- Of all things, Green Lantern got a reference that wasn't making fun of the movie. "Like a testicle with teeth" was Guy Gardner's line when describing what Zilius Zox looked like after taking leadership of the Red Lanterns. (Red Lanterns #22)
Mythology Gag / Deadpool (2016)