Sparkling wines are produced through a second fermentation of a base wine, whose composition strongly determines the final product quality. Yeast selection for primary fermentation is therefore crucial, as it influences both fermentation performance and sensory attributes. In this study, 156 yeasts were isolated from spontaneous fermentations of Vitis vinifera cv. Torrontés sanjuanino from Paraje de Hilario (1550 m.a.s.l., San Juan, Argentina), aiming to select autochthonous strains with oenological potential for sparkling base wine production. Isolates were phenotypically characterized and molecularly identified by sequencing the D1/D2 domain of the 26S rDNA. A total of 44 Saccharomyces cerevisiae and 60 non-conventional yeasts, mainly Hanseniaspora uvarum, were identified. Based on relevant oenological traits, two S. cerevisiae (M138M, F172M) and two H. uvarum (Mi14M, C135MJ) strains were selected. Laboratory and pilot-scale co-inoculation trials showed that the Mi14M/M138M (50:50) combination exhibited stable fermentation kinetics, low acetic acid production (0.44 g/L), high glycerol levels (7.1 g/L), and suitable pH (3.08) and ethanol content (11.2% v/v). Despite higher residual sugars than the control, no technological issues were observed. These findings support co-inoculation of autochthonous strains of S. cerevisiae and H. uvarum as a promising strategy to enhance wine quality and reinforce regional identity in sparkling base wines.