Habitat fragmentation poses a significant threat to the existence and reproduction of large carnivores, such as brown bears, as it affects the genetic connectivity of populations and, consequently, their long-term viability. Understanding the genetic makeup and dispersal patterns in areas where brown bears live is crucial for developing effective conservation plans and promoting human-brown bear coexistence. In this study, 214 hair samples were collected non-invasively from brown bears and were genetically analyzed using fifteen specific microsatellite loci to shed light on the genetic status and demography of a sub-population residing in Central Greece (Trikala-Meteora area). The broader Central and South Pindos regions have not been studied for over a decade; however, high levels of genetic diversity, a lack of heterozygosity deficiency, and no signs of inbreeding, along with an estimated effective population size (Ne = 99), confirm the good conservation status of this sub-population. Moreover, the existence of a natural and apparently functional corridor in Central Pindos was demonstrated by bear movements between the western and eastern parts of the project area, enhancing the sustainability of the population. These findings will contribute to future efforts to conserve natural corridors that enable functional connectivity among large areas of brown bear territories in Greece.