The paper focuses on the Pleistocene deposits in Perspektywiczna Cave, southern Poland, related to cave hyena (Crocuta crocuta). We used direct radiocarbon dating of hyena fossils supported by genetic and stable isotope analyses to infer the paleobiology of this population. Radiocarbon dating of 19 hyena remains suggests long inhabitation of the region during early MIS 3, around 50–34 ky cal BP. The youngest among our dates, 34,355–33,725 cal BP (1σ, combined of two dates for the same specimen) points out the latest appearance of a cave hyena north to Carpathians. Beside this long period of occupation, the Perspektywiczna Cave hyenas stayed ecologically stable, but their genetic structure changed. Two mtDNA haplogroups were present, one typical for other Late Pleistocene European populations and the other one known so far only from recent African populations.