The effect of temperature shock on androgenic ability was studied in a large number of different plant species. The optimal temperature and duration of pre-treatment, which will stimulate the androgenesis, is different for each plant species. Androgenic response in sunflower is generally low due to genotype and explant specificity and very low level of plant regeneration from haploid tissues. However, cold pre-treatment was found to have positive effect on callus and embryo and shoot regeneration from anthers. In our work, we have studied the effect on cold pre-treatment on callus induction and regeneration from F3 sunflower genotypes created at Institute of Field and Vegetable Crops, Novi Sad, Serbia (IFVCNS). One half of the anthers of each genotype was submitted to a cold pre-treatment, after which they were placed on KM and KM-R cultivation media and incubated. The other half of anthers of each genotype was incubated on the same cultivation media, but without the pre-treatment. After ten days of incubation in the dark, the anthers, which were not submitted to the cold pre-treatment, showed better callusogenesis than the anthers which were exposed to the cold, on both media. However, after the first subculture and moving to the light chamber, the anthers which were submitted to the cold pre-treatment showed a significant increase in callusogenesis percentage on both media. Similar to the results obtained by other authors, in our experiment genotype had a significant effect on the regeneration capacity. The results obtained in this experiment confirm the positive effect of cold pre-treatment on callusogenesis in tested sunflower genotypes. The differences in the percentage of callusogenesis among the genotypes were quite expected and only confirm the importance of optimizing the culture conditions for increased regeneration efficiency, less dependent on genotype.