The Dutchman picked up a hamstring strain during Sunday's El Clasico defeat to Real Madrid and is now facing one to two weeks on the sidelines, where he joins long-term victim Martin Braithwaite.
Riqui Puig and Nico Gonzalez are alternative options for Ronald Koeman in the middle of the park, but Roberto was De Jong's replacement at the weekend and should link up with Gavi and Sergio Busquets in the engine room.
Ansu Fati is also expected to be fine for Wednesday's game after shaking off his own injury scare, but Koeman may still be reluctant to throw the 18-year-old into the first XI twice in the space of a few days after his lengthy layoff.
As a result, a spot could open up for Sergio Aguero to make his first start for the club after netting his first Barca goal in El Clasico, with Memphis Depay and Sergino Dest providing support to the Argentine.
Barcelona possible starting lineup: Ter Stegen; Mingueza, Pique, Garcia, Alba; Roberto, Busquets, Gavi; Dest, Aguero, Depay