Murillo began his art studies in Seville under Juan del Castillo, who was a relative of his mother (Murillos uncle, Antonio Pérez, was also a painter). His first works were influenced by Zurbarán, Jusepe de Ribera and Alonzo Cano, and he shared their strongly realist approach. The great commercial importance of Seville at the time ensured that he was subject to artistic influences from other regions. He became familiar with Flemish painting and the "Treatise on Sacred Images" of Molanus (Ian van der Meulen or Molano). As his painting developed, his more important works evolved towards the polished style that suited the bourgeois and aristocratic tastes of the time, demonstrated especially in his Roman Catholic religious works.
In 1642, at the age of 26, he moved to Madrid, where he most likely became familiar with the work of Velázquez, and would have seen the work of Venetian and Flemish masters in the royal collections; the rich colors and softly modeled forms of his subsequent work suggest these influences. In 1645 he returned to Seville and married Beatriz Cabrera y Villalobos, with whom he eventually had eleven children.