Vittoria was betrothed to her cousin Ferdinando in 1623, married in 1634. While she was the potential heir to the duchy of Urbino when her grandfather, the duke Francesco Maria II della Rovere died, however, the duchy was annexed to the Papal States by Pope Urban VIII. The Medici however obtained in 1631 the rich art collections of the family, now in the Uffizi and Palazzo Pitti.
After two children who died early, disputes arose about the education of their son Cosimo (born in 1642), whom Ferdinando wanted to be educated in a modern secular fashion. In the end, Vittoria forced a strict Catholic education on the boy. Another child, Francesco Maria, was born in 1660.
Ferdinando and Vittoria had, at best, an unhappy marriage and lived separately for many years. In old age she made long stays in the convent of the Montalve known as La Quiete, as well as in the Villa del Poggio Imperiale. She died in Pisa in 1694.