The League Two side revealed in March that they would not be extending Lester's deal beyond the end of this season.
The 37-year-old, who joined Chesterfield from Nottingham Forest in 2007, will captain the side for the game against Exeter.
Lester told the club's official website: "I've decided to retire from football in the summer, which has been a tough decision. I've got some opportunities coming up to observe coaching around the world.
"I'll get an insight into some international sports psychologists, using their expertise. I'm also doing some coaching in America.
"I've got a year before my son starts school, which is why I've decided to retire and make the most of the opportunities that are there."
Lester has scored 83 goals in 196 league appearances for Chesterfield.