This chapter outlines the results of stable isotope studies of the prehistoric populations of Ukraine across the earlier to middle Holocene, ca. 10,000–3500 cal BC. The data are consistent with the continued exploitation of fisher-hunter-forager subsistence strategies across the periods studied with variations occurring in the relative amount of freshwater proteins consumed across the Epipalaeolithic through to Eneolithic periods. During the Neolithic and later periods there is a clear increase in the frequency of domesticates in zooarchaeological assemblages, indicative of an increasing emphasis on pastoralism and animal husbandry. However, the visibility of domestic fauna in dietary isotope studies is difficult to determine due to the paucity of faunal remains available for analysis. The key exception to the dominant subsistence trends appears to relate to the Trypillia farming culture, where agro-pastoralism is evidenced, and in this context isotope data from the site of Verteba Cave in western Ukraine is discussed. The majority of isotope data considered here are obtained from the cemeteries located in the Dnieper River valley, particularly those focused on the rapids, and its tributaries.