The Arsenal man has failed to recover from a knock sustained against Greece and misses the Group A decider, while Tyton keeps his place in goal for the co-hosts
An Achilles problem forced the 31-year-old captain off during the 2-1 victory, which gives Michal Bilek's side the opportunity to qualify for the last eight if they defeat Poland on Saturday evening. Viktoria Pilzen's Daniel Kolar will take Rosicky's place in the starting XI, as he did against the Greeks.
For Poland, meanwhile, coach Franciszek Smuda keeps faith with Przsemyslaw Tyton after the keeper impressed against Russia. The PSV man deputised for Wojciech Szczesny following his red card in the Group A curtain-raiser against Greece and although the Arsenal shot-stopper is available once more having completed his suspension, Tyton will start a game in which Poland need at least a draw to ensure their Euro 2012 campaign does not end at the first hurdle.
Tyton's inclusion means the Poles field an unchanged starting line-up, with Robert Lewandowski once more leading the line for the host nation.
Czech Republic (4-2-3-1): Cech; Gebre Selassie, Sivok, Kadlec, Limbersky; Hubschman, Plasil; Jiracek, Kolar, Pilar; Baros
Substitutes: Lastuvka, Drobny, Suchy, Hubnik, Rajtoral, Petrzela, Rekez, Rosicky.
Poland (4-3-3): Tyton; Piszczek, Wasilewski, Perquis, Boenisch; Murawski, Polanski, Dudka; Blaszczykowski, Lewandowski, Obraniak.
Substitutes: Szczesny, Sandomierski, Wojkowiak, Kaminski, Wawrzyniak, Rybus, Matuszczyk, Mierzejewski, Wolski, Grosicki, Sobiech, Brozek.