The effect of explant type (immature vs. mature embryos) and two auxin types (2,4-dichlorophenoxyacetic acid vs. Dicamba) on the callogenesis and plant regeneration ability of 26 wheat cultivars was studied. In general, the callus induction, plant regeneration and shoot formation frequencies were higher in mature embryo-derived cultures as compared to immature ones on media originally developed for mature wheat embryo cultures. In both culture types, the auxin Dicamba was found to be more efficient, especially when mature embryos were cultured. The separation of means using Duncan’s multiple range test revealed the best in vitro response, in terms of the frequency of callus regeneration, in the cultivar Astella for both immature and mature embryo cultures. This cultivar gave very promising results, suggesting that it could be used in the future for further tissue culture investigations and as a donor material for genetic transformation experiments in wheat. Correlation analyses revealed significant similarities between the evaluated parameters within each group (immature and mature embryo-derived cultures). However, there were no significant correlations between these two groups for most of the parameters. This suggests that the mechanism of plant regeneration in the two in vitro regeneration systems (mature vs. immature embryo culture) may be different enough to hamper the development of an optimal plant regeneration protocol for use in both systems.