Both teams' fate is sealed before the final round of northern section Group D fixtures, so only pride will be at stake when they lock horns at Holker Street.
Match preview
Without a victory in seven matches across all competitions, Barrow's winless streak began with a 3-0 EFL Trophy loss to Fleetwood Town last month, confirming their elimination even before the final matchday.
The Bluebirds had previously lost their opener to Bolton Wanderers, so there is no chance of securing a top-two finish in Group D this week, regardless of both results.
Since losing to Fleetwood, they have accrued just three points from a possible 15 throughout five League Two fixtures, suffering back-to-back defeats before holding Notts County and Bromley to 1-1 draws at the end of October.
Then, on Saturday, Stephen Clemence's side were forced to settle for the same result against Colchester, who struck in the second minute of stoppage time to earn a point at Holker Street.
Barrow were denied wins in both previous league games due to conceding last-gasp penalties, and it was another bitter pill to swallow for a side that have now slipped down into mid-table.
Clemence - whose team also crashed out of the FA Cup earlier this month - has been serving a three-match touchline ban after his red card at Bromley, so he will be keen to make his presence felt from the dugout and finally stop the rot against Aston Villa Under-21s.
Villa's Young Lions lead Group D by one point after edging a five-goal contest in their first fixture against Fleetwood - when senior star Emi Buendia completed his road to recovery by scoring twice on his first competitive start since May 2023 - then beating Bolton on penalties.
It has been a less fruitful season for Josep Gombau's boys in Premier League 2, though: after conceding three times in the first half-hour of last week's loss to Leicester City, they occupy 21st place in a 26-team table.
Some of the Under-21 squad were then involved in Villa's recent 6-2 UEFA Youth League victory over Club Brugge, having previously beaten Bologna.
Now, they have another chance to tackle full-time professionals in the EFL Trophy, before looking ahead to the knockout rounds, which will kick off in December.
Barrow EFL Trophy form:
L
L
Barrow form (all competitions):
L
L
D
D
L
D
Aston Villa Under-21s EFL Trophy form:
W
W
Aston Villa Under-21s form (all competitions):
W
L
L
W
L
L
Team News
Ahead of an important league clash with AFC Wimbledon, it is likely that Stephen Clemence will rest a whole host of regulars - particularly as Tuesday's game is effectively a dead rubber.
Connor Mahoney, Elliot Newby and several others may be left on the bench, with fringe players offered an opportunity instead.
Barrow lost Kian Spence to injury during their weekend draw with Colchester, so his inclusion is also in doubt.
Meanwhile, Aston Villa Under-21s will still field their strongest XI despite already sealing qualification, as Josep Gombau seeks to give more experience to his young squad.
Top scorer Ben Broggio and Kobei Moore - son of former Villa striker Stefan Moore - should both feature in the final third, perhaps joining 18-year-old winger Kadan Young, who has played twice for the senior side.
Barrow possible starting lineup:
Daniels; Eccleston, Vassell, Feely, Tiensa; Campbell, Foley; Garner, Telford, Kouyate; Dallas
Aston Villa Under-21s possible starting lineup:
Proctor; Rowe, Swinkels, Amundsen-Day, Patterson; Morland, Alcock, Jimoh-Aloba; Young, Moore, Broggio
We say: Barrow 1-2 Aston Villa Under-21s
While Barrow will boast more first-team nous and experience, their motivation will be relatively low given the circumstances.