Both finished second in their domestic league last season, and they are now just two steps away from the new-look group stage.
Match preview
Having been pipped to the Czech title by city rivals Sparta Prague for the second successive season, Slavia Prague's consolation prize was a place in this year's Champions League qualifiers, and victory in their opening tie would take them straight into the playoffs.
Following a successful pre-season, which featured four wins from five, Jindrich Trpisovsky's side have been back in competitive action since late last month, when they kicked off their league campaign with a disappointing 0-0 draw at Slovacko.
Subsequent wins over Slovan Liberec and Dynamo Ceske Budejovice have since lifted Slavia to their familiar place behind Sparta in the very early standings, and they have yet to concede a single goal across their first three matches.
The Cervenobili now make this season's European bow at Fortuna Arena, where in four Europa League contests last term they posted three wins and one loss, memorably beating Roma 2-0 in November.
As a result, they finished first in Group G with 15 points, and a 6-0 victory over Sheriff Tiraspol was the club's biggest in UEFA competition outside of qualifying - they also became the first Czech team to top their group.
Ultimately exiting to AC Milan in the last 16, losing 7-3 on aggregate, ended Slavia's dream of reaching a first continental final, but now their main aim is to dine at Europe's top table.
Union Saint-Gilloise take their place in qualifying after topping Belgium's regular-season table for a second time in three years, though they again finished runners-up - this time behind Club Brugge - in the final league standings.
Les Unionistes - who were in the second tier as recently as 2021, before remarkably finishing second in their top-flight comeback campaign one year later - fell just short of claiming the crown for a third straight season.
Nonetheless, they did finally end an almost 90-year wait for a major trophy by claiming the Belgian Cup, and new boss Sebastien Pocognoli has since made a successful start by winning the Super Cup against Club Brugge, with a 2-1 triumph at Jan Breydel Stadium.
After then dropping points away to newly-promoted Dender in a goalless opening-day draw, Union recorded their first Pro League win of 2024-25 with Friday's 3-1 defeat of Beerschot.
Last autumn, the Brussels club finished third in a Europa League group also featuring Liverpool, Toulouse and LASK, despite notably beating the Premier League side 2-1, and after battling past Eintracht Frankfurt in the Conference League playoffs, they ultimately lost to Fenerbahce in the last 16.
Along the way, they registered two wins, one draw and two losses in five games on their travels, and they now head for Prague seeking a positive result to take into next week's second leg.
Slavia Prague form (all competitions):
Union SG form (all competitions):
Team News
While Euro 2024 joint-Golden Boot winner Ivan Schranz and Slavia Prague's four-goal top scorer in last season's Europa League, Mojmir Chytil, are both available to start on Wednesday, the hosts may again be without their main marksman.
Vaclav Jurecka has topped the Czech First League's scoring charts for the past two years, netting 19 times last term, but he is yet to play a single minute this season due to a contract dispute.
Furthermore, Norwegian midfielder Conrad Wallem has been subject of strong interest from Preston North End, but he should still feature in the squad and could join Liberia captain Oscar Dorley in the hosts' engine room.
Meanwhile, Cameron Puertas was the leading scorer in Union SG's last European campaign, scoring three goals in 2023-24, and after finding the net again on Friday, he is set to start in support of Dennis Eckert up front.
The visitors will be without two central defenders on Wednesday, as English pair Ross Sykes and Christian Burgess have been ruled out by injury; Madagascar midfielder Loic Lapoussin is out in the cold following his recent transfer request.
Slavia Prague possible starting lineup:
Kinsky; Holes, Ogbu, Boril; Masopust, Wallem, Dorley, Diouf; Schranz, Chytil, Provod
Union SG possible starting lineup:
Moris; Mac Allister, Machida, Leysen; Castro-Montes, Ait El-Hadj, Sadiki, Puertas, Teklab; Kabangu, Eckert
We say: Slavia Prague 2-1 Union SG
After positive pre-seasons and good starts to their respective domestic campaigns, both teams are tuned up for this important first-leg contest.