The fifth of the seven children of Eugène de Beauharnais (1781–1824), Duke of Leuchtenberg, and his wife, Princess Augusta of Bavaria (1788–1851), Théodolinde was born in Mantua, Italy, and presumably named for Theudelinda, a 6th-century queen of the Lombards.

She had two brothers (Auguste and Maximilian) and three surviving sisters (Joséphine, Eugénie, and Amélie).

Joséphine de Beauharnais, the first wife of Napoléon Bonaparte and former Empress of France, was her paternal grandmother.

She died after a short illness on the morning of 1 April 1857 in Stuttgart, Germany, and was buried in the family vault at Ludwigsburg Palace, with her heart buried at the Hauskapelle of the palace in Munich.

He was also a cousin of Princess Catharina of Württemberg (1783–1835), who became the second wife of Jérôme Bonaparte, Napoleon's youngest brother, in 1807.