A number of senior players will be involved in Tuesday night's game at Sheffield FC (7pm).
A Spireites XI will make the short trip to Dronfield for the match, which will raise money for St Luke's Hospice, at the request of the family of former Chesterfield player Kevin Austin, who passed away recently at the age of 45.
Assistant-manager Adrian Whitbread said: "We will be taking a mixture of senior players, young pros and second-year scholars.
"Robbie Weir, Shwan Jalal, Michael Nelson and Kyel Reid are all set to feature. Younger players like Ify Ofoegbu, Charlie Wakefield and Jamie Sharman, who recently signed his first professional contract, will also be involved."
Admission will cost £5 for adults and £3 for concessions (adults in receipt of state pension/benefits, students and young persons aged under 18).