Cech missed a 2-2 away draw against Scotland in the previous Sept. 3 qualifier after injuring a knee ligament in training with Chelsea. Rosicky, meanwhile, had recovered from a knee injury in time to be fielded as a substitute in his Arsenal's 3-0 win over Bolton on Saturday.
Czech Republic coach Michal Bilek will be without midfielder Jaroslav Plasil and striker Jan Rezek against Spain in Prague on Oct. 7, with both suspended. They will be available against Lithuania four days later.
Spain has qualified from Group I while the Czechs are battling with Scotland for a playoff spot.
Goalkeepers: Petr Cech (Chelsea), Jaroslav Drobny (Hamburg), Jan Lastuvka (Dnipropetrovsk).
Defenders: Roman Hubnik (Hertha Berlin), Michal Kadlec (Bayer Leverkusen), Zdenek Pospech (Mainz), Daniel Pudil (Genk), Jan Rajnoch (Ankaragucu), Tomas Sivok (Besiktas), Theodor Gebre Selassie (Liberec).
Midfielders: Tomas Hubschman (Shakhtar Donetsk), Petr Jiracek (Plzen), Daniel Kolar (Plzen), Marek Matejovsky (Sparta Prague), Milan Petrzela (Plzen), Vaclav Pilar (Plzen), Jaroslav Plasil (Bordeaux), Tomas Rosicky (Arsenal), Kamil Vacek (Sparta Prague).
Forwards: Milan Baros (Galatasaray), David Lafata (Jablonec), Tomas Pekhart (Nuremberg), Jan Rezek (Anarthosis Famagusta).