Jurgen Klopp's men made it all the way to the final of the competition last term before being beaten 2-0 after extra-time by Bayern Munich.
Dortmund head into their weekend fixture against third-tier Kickers on the back of a more positive meeting with their Bavarian rivals, having inflicted a 2-0 defeat on Pep Guardiola's side in the DFL Supercup earlier this week.
Kickers are already four games into their league campaign, and sit sixth in the 3. Liga with seven points so far.
Although Kickers will officially be the home side, the match will actually take place at the Mercedes-Benz Arena rather than their own GAZI-Stadion auf der Waldau.
The Stuttgart outfit face a huge task on Saturday, as three-time Pokal winners Dortmund have only lost two of their last 16 matches in the competition - both to Bayern.
With the start of the new Bundesliga campaign coming next week, Klopp could opt to rest some of his big guns, while several players will miss out through injury.
Oliver Kirch (foot) is expected to be sidelined, while Marcel Schmelzer is out with an abductor injury and defender Sokratis Papastathopoulos is also a doubt after picking up a knock on Wednesday.