Asensio, 21, made his Spain debut in the Euro 2016 warm-up match against Bosnia-Herzegovina last May but has not appeared for the national team since.
Real Sociedad midfielder Asier Illarramendi is also included, as is Athletic Bilbao goalkeeper Kepa. Manchester United's Juan Mata, who has recently returned to action following groin surgery, again misses out.
Spain take on Colombia in a friendly on June 7 before a World Cup qualifying clash with Macedonia four days later.
The Real Madrid players named in the squad, who include Asensio, Dani Carvajal, Sergio Ramos, Nacho and Alvaro Morata, are expected to miss the Colombia match due to the Champions League final on June 3.
Squad in full:
Goalkeepers: David de Gea (Manchester United), Kepa (Athletic Bilbao), Pepe Reina (Napoli)
Defenders: Jordi Alba (Barcelona), Cesar Azpilicueta (Chelsea), Dani Carvajal (Real Madrid), Nacho (Real Madrid), Nacho Monreal (Arsenal), Gerard Pique (Barcelona), Sergio Ramos (Real Madrid), Sergi Roberto (Barcelona)
Midfielders: Thiago Alcantara (Bayern Munich), Marco Asensio (Real Madrid), Sergio Busquets (Barcelona), Asier Illarramendi (Real Sociedad), Andres Iniesta (Barcelona), Isco (Real Madrid), Koke (Atletico Madrid), Pedro (Chelsea), David Silva (Manchester City), Vitolo (Sevilla)