On her paternal side, she was the granddaughter of the first Margrave of Mantua, Gianfrancesco Gonzaga, and his wife Paola Malatesta da Rimini.

[3] Finally, at the suggestion of Albrecht III Achilles, Elector of Brandenburg (Barbara's great-uncle), was arranged her marriage with Eberhard V, Count of Württemberg-Urach.

Barbara joined her husband a few months later, escorted by horse guards led by her brother Rodolfo Gonzaga.

During 4–7 July 1474 in the Urach Castle (who was renovated and enlarged for this occasion) and in the presence of numerous high-ranking guests (among them were the Bishops of Konstanz, Speyer and Augsburg, Elector Albrecht III Achilles, Philip, Count Palatine of the Rhine, Otto II, Count Palatine of Mosbach-Neumarkt and Margrave Charles I of Baden-Baden), celebrations were held on the occasion of the arrival of the new Countess.

His educated wife introduced him to Italian art, music, and humanist philosophy; she was instrumental in founding the University of Tübingen, supporting her husband in the implementation of this project.

[3][6] On 2 August 1475 Barbara gave birth a daughter, who was named after her; unfortunately, the child lived only two months, dying on 15 October and buried in the charterhouse of Güterstein.

The inability to give birth to an heir to her husband was probably the reason who accentuated the innate tendency of Barbara to depression.

The stained glass window depicting the Duchess of Württemberg in the University of Tübingen was made in modern times and is the author's fantasy.