The Chauvet-Pont d'Arc and Deux-Ouvertures caves, located along the Ardèche River (France), contain abundant remains of the extinct cave bear (Ursus spelaeus). Because they also display a variety of Palaeolithic anthropogenic evidences, such as the earliest charcoal drawings recorded to date (Chauvet-Pont d'Arc), and delicate engravings (Deux-Ouvertures), they offer the opportunity of studying the interaction between animals and human beings during a key period for Pleistocene species extinctions. We characterized cave bear specimens from these two sites by radiocarbon dating, stable isotopes, and mitochondrial DNA analysis. In Chauvet-Pont d'Arc, we obtained radiocarbon ages that ranged between 29,000 and 37,300 years before present (BP). The Deux-Ouvertures cave bear specimens clustered to the bottom of this time frame, returning radiocarbon ages of 27,440–30,220 years BP. Cave bear nitrogen isotope values were all compatible with a vegetarian diet. Mitochondrial DNA analysis, carried out on a highly variable domain of the control region, evidenced only two cave bear haplotypes, including a new haplotype, and a common one which largely predominated. We detected both haplotypes in Chauvet-Pont d'Arc, but only recorded the predominant one in the Deux-Ouvertures Cave. Our data put forward the surprising observation that cave bears inhabited Ardèche over a short period of time, from about 37,000 to 27,400 years BP. They were notably present during the first (Aurignacian) phase of human intrusions in Chauvet-Pont d'Arc, 30,000–32,000 years BP. This points to the possible competition for cave sites, presumably on a seasonal scale considering the cave bear habit for hibernation. During this time period, the small number of haplotypes is at variance with the extensive genetic diversity reported elsewhere for much more ancient specimens.Highlights : ► We compare extinct cave bears from Chauvet and Deux-Ouvertures caves (France). ► Mitochondrial DNA analysis unveils a new cave bear haplotype. ► Comparison between the two caves shows strong genetic homogeneity. ► Radiocarbon dating reveals that the specimens date back to 27–37 ky BP