This study focuses on the spatial differentiation characteristics and multidimensional influencing mechanisms of homestays in Hunan Province. Based on data from 6,508 homestays collected from OTA platforms such as Meituan and Ctrip before March 2024, the study uses ArcGIS software combined with spatial analysis methods such as the Nearest Neighbor Index, kernel density estimation, and Lorenz curve. The findings reveal that the spatial distribution of homestays in Hunan presents an overall pattern of “dense in the east and sparse in the west, with agglomeration differentiation.” Key factors influencing the spatial layout of homestays include university agglomeration, transportation accessibility, and the endowment of tourism resources. Notably, the “education-driven” agglomeration effect centered around higher education institutions is a unique spatial feature in Hunan, differentiating it from other regions where transportation or tourist attractions serve as the core drivers of spatial layout. Furthermore, this study identifies the collaborative impact of multiple factors at the regional scale and the dynamic evolution of spatial patterns, shedding light on the inherent logic of cultural-tourism spatial reconstruction. The findings provide significant empirical support and decision-making references for Hunan’s efforts to promote a “university-tourist attraction-transportation” three-dimensional collaborative governance system and formulate cultural tourism integration policies.