Bartol was born on February 24, 1903 in San Giovanni (Slovene: Sveti Ivan), a suburb of the Austro-Hungarian city of Trieste (now in Italy), as the third child of seven children of a middle class Slovene family. His father Gregor Bartol, was a post office clerk, and his mother Marica Bartol Nadlišek was a teacher, a renowned editor and feminist author. Vladimir's parents offered their children extensive education. His mother introduced him to painting, his father to biology. In his autobiographical short stories, Bartol described himself as an oversensitive and slightly odd child with a rich fantasy life. He was interested in many things: biology and philosophy, psychology, art, as well as theatre and literature. As a scientist, he collected and researched butterflies.

Vladimir Bartol began his elementary and secondary schooling in Trieste and concluded it in Ljubljana, where he enrolled at the University of Ljubljana to study biology and philosophy. In Ljubljana, he met the young Slovene philosopher Klement Jug who introduced him to the works of Friedrich Nietzsche. Bartol also gave special attention to the works of Sigmund Freud. He graduated in 1925 and continued his studies at Sorbonne in Paris (1926–1927), for which he obtained a scholarship. In 1928 he served the army in Petrovaradin (now in Serbia). From 1933 to 1934, he lived in Belgrade, where he edited the Slovenian Belgrade Weekly. Afterward, he returned to Ljubljana where he worked as a freelance writer until 1941. During World War II he actively participated in the resistance movement. After the war he moved to his hometown Trieste, where he spent an entire decade, from 1946 to 1956. Later he was elected to the Slovenian Academy of Sciences And Arts as an associate member, moved to Ljubljana and continued to work for the Academy until his death on September 12, 1967. He is buried in the Žale cemetery in Ljubljana.