These two sides last met in 2018 when they were part of the same UEFA Nations League group, and it was a closely-fought contest with Ukraine coming out as 1-0 winners to top that group.
Match preview
Jaroslav Silhavy's side began this international week with a positive 1-0 win over Belarus last Thursday, thanks to a 34th-minute strike from Antonin Barak.
However, any hopes of topping Group E in the World Cup qualifiers started to diminish after Czech Republic suffered a heavy defeat when they travelled to face group favourites Belgium on the weekend.
Romelu Lukaku scored within eight minutes to put the home side ahead, with Eden Hazard doubling Belgium's advantage just before half time, and Alexis Saelemaekers capped off a dominant performance over Czech Republic.
Those two results have left Silhavy's side in second place going into their final three fixtures, but Wales sit just one point behind and have two games in hand.
Czech Republic face Wales at the beginning of October in what looks to be a decisive game for second place in the group, so they will want to go into that game with a victory against Ukraine under their belt for confidence.
Ukraine have had a mixed set of results this September picking up two draws, but one point against Kazakhstan will feel like a missed opportunity to try and catch Finland in second place.
Danylo Sikan thought he had won the game for Ukraine last Wednesday when he scored in the 93rd minute, but Kazakhstan's Ruslan Valiullin denied the away side all three points with a strike in the 95th minute.
Table toppers France visited Kiev for their second fixture this month, and a 1-1 draw against the World Champions will be seen in a more positive light.
Mykola Shaparenko put the home side 1-0 up heading into half time, but Anthony Martial equalised for France just after the restart meaning that the points were shared, and that has left Ukraine level on points with Finland, who have two games in hand on Ukraine.
A tied scoreline involving Oleksandr Petrakov's side in this World Cup qualifying campaign does not come as a surprise, with the team having drawn all five of their fixtures so far.
Ukraine will be looking to emulate a more positive result in their friendly against Czech Republic on Wednesday, like they managed in their last friendly before the Euros, beating Cyprus 4-0 at home.
Czech Republic International Friendlies form:
Czech Republic form (all competitions):
Ukraine International Friendlies form:
Ukraine form (all competitions):
Team News
A friendly fixtures potentially gives the opportunity to both managers to give the fringe players a chance to start for their country and earn a place in the team for next month's World Cup qualifiers.
That means players such as Michal Sadilek and Jaromir Zmrhal may be given the chance to play from the off on Wednesday, having either come on from the bench or not at all during September's fixtures.
Some key players Silhavy is unlikely to change include West Ham United duo Vladimir Coufal and Tomas Soucek, along with Bristol City centre-back Tomas Kalas.
Ukraine altered to a back five for their game against France on Saturday, but they are likely to play in a 4-4-1-1 formation as they did against Kazakhstan to start this international week of games.
Manchester City's Oleksandr Zinchenko is closing in on 50 caps for his country, and he is likely to play on Wednesday which would mean that he could achieve his half century of appearances by the end of 2021, if he features in all five Ukraine games left this year.
Czech Republic possible starting lineup:
Vaclik; Mateju, Holes, Kalas, Coufal; Soucek, Sadilek; Zmrhal, Barak, Hlozek; Dolezal
Ukraine possible starting lineup:
Pyatov; Mykolenko, Matviyenko, Zabarnyi, Karavaev; Sobol, Zinchenko, Sydorchuk, Yarmolenko; Buyalsky; Yaremchuk
We say: Czech Republic 2-1 Ukraine
Czech Republic will be determined to bounce back from their 3-0 defeat to Belgium and a friendly fixture provides them with a good opportunity to head into their World Cup qualifiers in October with another victory to their name.
Ukraine have struggled to turn their draws into three points throughout their World Cup qualifying campaign, and with Czech Republic having home advantage on Wednesday, it could make that a difficult task for Petrakov's side.