The draw for the Checkatrade Trophy has been made with Category One Academies drawn against sides from Leagues One and Two.
Last season's Premier League 2 champions Arsenal will compete in the competition for the first time and have been drawn in Group E of the Southern Section.
Chelsea, who reached the semi-finals of the 2017/18 competition, are in a group with Newport County, Plymouth Argyle and Swindon Town.
'A real challenge'
Wolverhampton Wanderers reached the third round when they last appeared in the competition in 2016/17 and they are in a group with holders Lincoln City.
Scott Sellars, Wolves' head of Academy player development, says it will be a valuable experience for his players.
"The last time we were in it I spoke very highly of the competition," he says. "I gave us something that we didn't have which was to play against men in front of crowd at a proper stadium.
"The players thrived in it, they loved it. It really challenged them."
Northern Section
Group A: Carlisle, Morecambe, Sunderland, Stoke U21s
Group B: Bury, Fleetwood, Rochdale, Leicester U21s
Group C: Accrington, Blackpool, Macclesfield, West Brom U21s
Group D: Crewe, Shrewsbury, Tranmere, Man City U21s
Group E: Burton, Port Vale, Walsall, Middlesbrough U21s
Group F: Barnsley, Bradford, Oldham, Everton U21s
Group G: Doncaster, Grimsby, Notts County, Newcastle U21s
Group H: Lincoln, Mansfield, Scunthorpe, Wolves U21s
Southern Section
Group A: Crawley, Gillingham, Portsmouth, Spurs U21s
Group B: Cambridge, Colchester, Southend, Southampton U21s
Group C: Newport, Plymouth, Swindon, Chelsea U21s
Group D: Bristol Rovers, Exeter, Yeovil, West Ham U21s
Group E: Cheltenham, Coventry, Forest Green Rovers, Arsenal U21s
Group F: Northampton, Oxford, Wycombe, Fulham U21s
Group G: AFC Wimbledon, Charlton, Stevenage, Swansea U21s
Group H: Luton, MK Dons, Peterborough, Brighton U21s