Six clubs, Arsenal, Liverpool, Manchester United, Manchester City, Tottenham Hotspur and Newcastle have rejected the chance to play Under-21 teams in the EFL Trophy
Arsenal, Liverpool, Manchester United, Manchester City, Tottenham Hotspur and Newcastle have rejected the chance to play Under-21 teams in the EFL Trophy it was confirmed on the day the draw for the group games was announced.
Sixteen Category One academies were invited to play against League One and League Two clubs as part of a one-season, 64-team competition trial.
Premier League champions Leicester and Chelsea have accepted the invitation.
"If it's a success it will continue. If not we'll review and see where we go," said EFL chief executive Shaun Harvey.
National League champions Cheltenham return to the EFL Trophy with Northern Group A matches against Everton, Bolton and Blackpool - places that are a combined 453 miles away from the Gloucestershire town
Leicester and Chelsea are joined by Southampton, West Ham United, Stoke City, Everton, Swansea City, West Bromwich Albion, Sunderland and Middlesbrough in accepting the initial invitation to join the competition.
Six Championship academy teams will take the places of the rejected teams.
Norwich City, Brighton, Derby, Wolves, Blackburn and Reading have all put teams forward to play in the competition in 2016-17, which has undergone a revamp from the knockout stages of previous years.
Sixteen groups of four teams will each contain an academy side, with the top two from each qualifying for the first knockout round of 32 teams.
Category One academy teams will have to start six under-21 players in the competition, but the others can be of any age.
Barnsley beat Oxford United 3-2 at Wembley in front of a crowd of more than 59,000 in last season's final.
League Two play-off winners AFC Wimbledon, who voted against changes to the competition, are in the same group as Newport, Swansea and Plymouth
Northern Group A: Everton, Bolton, Cheltenham, Blackpool.
Northern Group B: Wolves, Chesterfield, Crewe, Accrington.
Northern Group C: Stoke, Bradford, Morecambe, Bury.
Northern Group D: Blackburn, Fleetwood, Carlisle, Oldham.
Northern Group E: Derby, Port Vale, Doncaster, Mansfield.
Northern Group F: Sunderland, Rochdale, Hartlepool, Notts County.
Northern Group G: Middlesbrough, Scunthorpe, Cambridge, Shrewsbury.
Northern Group H: Leicester, Sheffield United, Grimsby, Walsall.
Southern Group A: Reading, Bristol Rovers, Portsmouth, Yeovil.
Southern Group B: Swansea, AFC Wimbledon, Newport, Plymouth.
Southern Group C: Chelsea, Swindon, Exeter, Oxford.
Southern Group D: West Ham, Coventry, Wycombe, Northampton.
Southern Group E: Southampton, Charlton, Colchester, Crawley.
Southern Group F: Norwich, Peterborough, Barnet, MK Dons.
Southern Group G: Brighton, Southend, Stevenage, Leyton Orient.
Southern Group H: West Bromwich Albion, Millwall, Luton, Gillingham.