The reserves came from behind to beat Tranmere 4-2 on Tuesday afternoon at the Proact Stadium.
The visitors took a 13th minute lead when Joe Conchie produced a near-post finish after getting on the end of a low cross from Joe Thompson.
Tranmere failed to make the most of some good chances before Kieron Cadogan, who was playing on trial, equalised in the 26th minute after Jack Lester back-heeled the ball into his path.
Craig Clay (39) put Chesterfield in front after being set up by Mark Randall.
Jonathan Wafula (75) added a third goal with a neat finish after receiving the ball from fellow substitute Jack Waddle.
Cole Stockton reduced the arrears a minute later with a near post finish and Tranmere threatened to level when Danny Harrison fired a shot on target but young keeper Cameron Mason produced a good save to keep the ball out.
Another substitute, Mark Fereday, got his name on the score-sheet in stoppage time, turning home the loose ball after the Tranmere defence failed to cut out Jacob Hazel’s low cross.
The performance pleased Mark Crossley who said: “There were plenty of good things in the game.
“I was really pleased with the effort from the senior players and the young players can learn from them. We want to bring the young players on and this experience will help them develop.”
It remains to be seen whether Cadogan, who is a free agent after being released by Crystal Palace, did enough to convince the management team that he is worth an extended trial.
“He got a goal and I thought he looked okay,” said Crossley. “I’ll sit down with Tommy (Wright) and we’ll have a chat about him.”