Abstract: The Central Andes are characterized by the early emergence of complex societies and a chequered yet continuous cultural tradition. However, at least for certain points of time in the cultural development, the overall cohesiveness of this ‘culture area’ has been called into question, favoring an alternative perspective that emphasizes the existence of several relatively independent nuclei of development on the North Coast, the southern Peruvian Highlands and the Titicaca basin, with distinct cultural expressions and political organization. Here, we engage archaeological evidence and its interpretation with newly emerging perspectives from linguistics and genetics (modern and ancient DNA), including new targeted genetic analysis, to add fresh evidence to the question of the internal structure and cohesiveness of the ancient Central Andes as a culture area. The double cultural/biological approach points at a North vs. South structure bisecting the Central Andes that becomes appreciable ~2,000 years ago; however, as the evidence from all three disciplines indicates, too, the spheres have remained connected and hence maintained an overall cohesiveness. Our analysis suggests that demographic population structure precedes the constitution of distinct cultural domains, a pattern which is to be verified in other chronological transects in South America and at a global scale.