The quantity and quality of seedlings in the nursery has an impact on the success of re-establishment and later growth. High germination rates enable a sufficient number of seedlings, and their quality is assessed using a number of parameters, including seedling height and root collar diameter. These parameters are influenced in some species by seed size, but the correlation between them is species-specific. The model species in this research was sweet chestnut (Castaneasativa Mill.), and seeds from 12 populations from two distinct biogeographical regions of Croatia were collected. We examined the influence of seed size on four parameters: germination rate, seedling height, root collar diameter and sturdiness quotient. Seed size has been shown to have a positive influence on both seedling height and root collar diameter, whereas no such correlation was noted for germination rate and sturdiness quotient. Significant differences in nut size and seedling growth parameters were found between the Mediterranean and continental populations, with higher values observed in the coastal Mediterranean populations. We concluded that seed origin and seed size have a significant impact on seedling growth and are important factors to consider when choosing seed material. Further nursery operations should consider seed origin and local environmental conditions when choosing seedlings for reforestation efforts and general forest operations.