Valentine is a ruthless combat medic and the sole survivor of the Last Hope, a specialized anti-Skullgirl strike team. She originally served Lab Zero as a spy and researcher before her entire unit was wiped out, leading her to reluctantly work for the Skullgirl out of self-preservation. She fights using surgical tools adapted for combat, including bone saws, scalpels, IV stands, and body bags, and relies on high-mobility movement to overwhelm opponents. Her special moves include throwing scalpels, sliding across the floor with a medical saw, and dropping chemical vials onto the stage to debuff enemies with poison, slower movement, or increased stun times. Visually, she is styled like a cold-blooded assassin wearing a nurse uniform decorated with medical crosses and a surgical mask.

Easter is a combat medic from the elite squad The Last Hope in Skullgirls, best known for carrying a heavy bodybag into battle. She serves alongside Valentine and Patty under Anti-Skullgirl Lab 0, and her legacy lives on through Valentine, who uses Easter's signature bodybag as a weapon in her standard moveset.

Christmas is the former strict and solemn leader of Last Hope, an elite black-ops combat medic unit from Lab 7 in Skullgirls. As Valentine's squad leader and former rival, she wields a large scalpel sword and throwing knives in battle, sharing a visual design modeled after a distinct pink-accented nurse uniform.

Hallow is a combat nurse and member of the Anti-Skullgirl Labs black-ops medical unit known as The Last Hope. ​She serves alongside her teammates—Christmas, Valentine, Easter, and Patty—and shares a particularly close bond with Valentine. Visually, she sports a dark nurse uniform styled after a Halloween aesthetic, complete with a bat-like emblem on her cap and multiple giant syringes strapped to her arms. In combat, she relies heavily on these oversized syringes filled with various chemical concoctions, referring to them playfully as her "Tricks" and "Treats". Personality-wise, Hallow is mischievous and playful with a slightly eccentric, eerie edge that occasionally creeps out her fellow operatives. Valentine utilizes Hallow's signature syringes as one of her special memento attacks during battle.