For the first time, the Turtles are each of a different species rather than being blood-brothers of the same species:
- Raphael is a snapping turtle.
- Leonardo is a red-eared slider turtle.
- Donatello is a softshell turtle.
- Micheangelo is a box turtle.
Rob Paulsen, who voiced Raphael in Teenage Mutant Ninja Turtles (1987) and Donatello in Teenage Mutant Ninja Turtles (2012), serves as the voice director for this series.
Series producers Ant Ward and Andy Suriano cite the fantasy film Big Trouble in Little China (1986) as an influence on the show: "Modern New York is a melting pot. Wouldn't it be great if we found a way to explore that melting pot of mythological creatures and fables of all New York?"
In nearly all TMNT media, Leonardo wields two ninjato (a medium-sized sword with a straight blade), which are often incorrectly called katana (longer sword with a curved blade). In this show, Leo wields only one sword which hasn't happened since the live-action TV series Ninja Turtles: The Next Mutation (1997).
Donatello's gear in this series (a electrified bo staff, a drone) is based on his gear from Teenage Mutant Ninja Turtles: Out of the Shadows (2016).