The Red Devils won this competition 18 months ago, providing Erik ten Hag with his first piece of silverware at the club, and they come into the tie on the back of a morale-boosting victory at the weekend.
Match preview
After being thrashed 3-0 by rivals Liverpool before the international break, Man United desperately needed to respond when they got the Premier League weekend underway early on Saturday.
At St Mary's, United duly obliged, but they were given a big scare in a slightly flattering 3-0 victory over Southampton.
Ten Hag's side struggled to deal with the Saints' wide players for the opening half hour, and Diogo Dalot's rash challenge saw them concede a penalty.
Andre Onana proved his worth to the side though, saving from Cameron Archer, and United never looked back, as Matthijs de Ligt and Marcus Rashford scored quick-fire goals to put the visitors two-up at the break.
From there on out, Man United controlled proceedings, and a late third goal from Alejandro Garnacho after Southampton had gone down to 10 men added some gloss to the scoreline.
Attention now turns to cup matters, as the 2023 winners look to reach the fourth round - the stage where they were knocked out last season.
Newcastle United came to Old Trafford and beat Ten Hag's men 3-0, and even though they would go on to win the FA Cup to make amends for that early elimination, a better performance this season is imperative.
While third-tier Barnsley should not strike much fear into the hosts, Man United have suffered five defeats to lower-league opposition in this competition since 2011.
That will give the visiting side belief, but they come into the encounter on the back of a potentially damaging defeat at the weekend.
After taking 10 points from their four league games prior, a 3-0 loss at Stevenage on Saturday was Barnsley's first in the league since the opening day.
That means Darrell Clarke's side have dropped out of the League One playoff places for the time being, but they are still well positioned after a respectable start.
With two defeats in the league, they are expected to suffer another here, especially considering their only other loss this season came against Man United's Under-21s in the EFL Trophy.
In fairness, Barnsley were largely changed for that one, and Clarke has gone stronger in the EFL Cup, beating Wigan Athletic on penalties in round one, before an impressive win over Sheffield United in the previous round.
This will be a first meeting between the sides since United knocked Barnsley out of this competition at the last 16 stage in 2009, but the visitors have enjoyed success over their upcoming hosts in recent memory, beating them in the FA Cup back in 1998.
Manchester United form (all competitions):
Barnsley EFL Cup form:
W
W
Barnsley form (all competitions):
L
D
W
W
W
L
Team News
Ten Hag is likely going to use this clash as an opportunity to give minutes to some of the fringe players in his squad, while defensive injuries could also force his hand.
De Ligt, Lisandro Martinez and Noussair Mazraoui all went off at the weekend, but the former has revealed it was only due to cramp.
Martinez and Mazraoui will be assessed ahead of this one though, meaning the likes of Harry Maguire and Jonny Evans, who played in the previous meeting back in 2009, are likely to start.
Manuel Ugarte could be set for his full debut after coming off the bench at Southampton, while Casemiro, Antony and Garnacho could all be promoted from the bench.
Barnsley's one confirmed absentee is former Premier League midfielder Conor Hourihane, as attackers Max Watters and Fabio Jalo both returned from injury at the weekend off the bench.
Defender Donovan Pines was taken off at half time in the defeat to Stevenage, but it is believed it was precautionary and he should be fine to start here.
Manchester United possible starting lineup:
Onana; Dalot; Maguire, Evans, Amass; Casemiro, Ugarte, Collyer; Antony, Zirkzee, Garnacho
Barnsley possible starting lineup:
Slonina; Cotter, De Gevigney, Pines, Earl, Lofthouse; Connell, Phillips; Keillor-Dunn, Jalo, Humphreys
We say: Manchester United 2-0 Barnsley
United are set to rotate heavily with important domestic and European games on the horizon, but even their second-string side should have enough to overcome third-tier opposition here.