Non-marine Oligocene ostracods from Tremembé Formation (Taubaté Basin, Southeast Brazil) are studied for the first time. The study reveals rich assemblages which are probably composed of many new taxa, two of which are described here. The sixteen ostracod species registered are classified in the genera Cypretta Vávra, Strandesia Stuhlmann, Potamocypris Brady, Heterocypris Claus, Eucypris Vávra, Herpetocypris Brady and Norman, Cytheridella Daday and Limnocythere Brady. Two new species of the latter are herein proposed: L. mandubi sp. nov. and L. katu sp. nov. The succession of ostracod assemblages along the studied core changes conspicuously in composition, abundance and preservation, and are characterized by the following associations: Herpetocypris-Cytheridella (lower), Limnocythere-Cypretta (middle) and Potamocypris-Heterocypris (upper). It is assumed that these associations represent different ecological phases of the paleolake Taubaté which is in accordance to previous stratigraphic and paleontological studies in the basin. The results from this pioneering taxonomic and paleoecological study on ostracods from Tremembé Formation reinforce the potential of these fossils for paleolimnological researches in Brazilian Cenozoic deposits.