Assistant-manager Tommy Wright, reserve-team coach Mark Crossley, development coach John Dungworth and kit man Kurt McDermott all departed in a backroom re-shuffle.
Manager Paul Cook said: "On behalf of the club, I'd like to thank all the staff for their efforts and the support they gave me. I wish them all the best for the future."
Wright was named as former manager John Sheridan's assistant in June 2009 after the pair worked together previously at Oldham Athletic. After helping the Spireites win the League Two title and the Johnstone's Paint Trophy, he had a spell as caretaker manager earlier this season following Sheridan's departure.
Crossley also followed Sheridan from Oldham in the summer of 2009. He assisted Wright during his time in caretaker charge and looked after the reserves with Dungworth.