Paraguay face a friendly encounter against Greece in Switzerland on Wednesday as part of their warm-up for the tournament in South Africa.
In the World Cup, Paraguay will face reigning champions Italy, New Zealand and Slovakia in Group F.
Also among those included were Borussia Dortmund striker and freshly nationalised Argentine Lucas Barrios.
Barrios, whose mother is Paraguayan, has enjoyed a fine season with Dortmund, as has club mate Nelson Haedo Valdez, who also made the 23.
"I can imagine Paraguay causing some surprises in South Africa," said Barrios, who has scored 18 goals in the Bundesliga this season.
Santa Cruz has had a stop-start season with Manchester City because of a calf injury playing 20 games and scoring only four times since joining the club in June for £18m.
Goalkeepers: Justo Villar (Valladolid/ESP), Aldo Bobadilla (Deportivo Independiente de Medellin/COL), Diego Barreto (Cerro Porteno).
Defenders: Claudio Morel Rodriguez (Boca Juniors/ARG), Denis Caniza (Leon/MEX), Paulo da Silva (Sunderland/ENG), Dario Veron (Pumas UNAM/MEX), Julio Cesar Caceres (Atletico Mineiro/BRA), Carlos Bonet (Olimpia), Aureliano Torres (San Lorenzo de Almagro/ARG), Antolin Alcaraz (FC Bruges/BEL).
Midfielders: Edgar Barreto (Atalanta/ITA), Cristian Riveros (Cruz Azul/MEX), Victor Caceres (Libertad), Enrique Vera (Liga Deportiva Universitaria de Quito/ECU), Jonathan Santana (Wolfsburg/GER), Nestor Ortigoza (Argentinos Juniors/ARG).
Forwards: Roque Santa Cruz (Manchester City/ENG), Nelson Haedo Valdez (Borussia Dortmund/GER), Oscar Cardozo (Benfica/POR), Edgar Benitez (Toluca/MEX), Lucas Barrios (Borussia Dortmund/GER), Rodolfo Gamarra (Libertad).
This page is best viewed in an up-to-date web browser with style sheets (CSS) enabled. While you will be able to view the content of this page in your current browser, you will not be able to get the full visual experience. Please consider upgrading your browser software or enabling style sheets (CSS) if you are able to do so.