First multi-individual Neanderthal mitogenomes from north of the Carpathians

Neanderthals of Central-Eastern Europe are well documented by a wealth of archaeological sites, but thus far they remain poorly represented by both fossil and genetic data.1,2,3 At Stajnia Cave (Poland), nine Neanderthal teeth have now been integrated into a single high-resolution study combining morphological assessment, radiocarbon dating, and complete mitochondrial (mt) genome sequencing. We report eight new mitogenomes, including from four never-before-analyzed teeth, that resolve a minimum of seven, and possibly eight individuals. Three of the specimens share identical mitochondrial DNA (mtDNA), indicating that they are either from the same or maternally related individuals. Molecular branch shortening estimates place all samples in marine isotopic stage (MIS) 5, with point estimates of ∼119,700-92,498 years ago, making this the oldest multi-individual Neanderthal genetic assemblage yet characterized in Central Europe. Other Neanderthals with similar haplotypes were present in southeastern France, Iberia, and the Caucasus, suggesting this mt lineage might have been widespread across Europe before being replaced with the mtDNA of the "late Neanderthal type." Our analysis of the Stajnia fossils positions Central-Eastern Europe not as a peripheral fringe, but as a pivotal area for tracing Neanderthal geographic distribution.