Wheat is the most extensively cultivated and extremely nutritious cereal crop in the world. Increasing wheat productivity is essential to closing the food gap in Egypt. Applying modern biotechnological plant breeding techniques could create new, highly productive cultivars. Tissue culture is a powerful tool that can be used to facilitate genetic transformation, induce genetic variation in plants, and create new and improved crop cultivars. The present study examines the in vitro callogenesis expression and regeneration capacity of wheat cultivars in controlled laboratory conditions. Seeds from ten modern Egyptian commercial cultivars (five durum and five bread wheat cultivars) were collected for in vitro studies. Two different explants (immature embryos and mature embryos) and different media supplemented with different plant growth regulators were used to test the best wheat callus formation protocol. Immature embryos showed the highest callus formation value on MS medium supplemented with 2 mg/l 2,4-dichlorophenoxyacetic acid, while mature embryos showed the highest callus formation value on MS medium supplemented with 3 mg/l 2.4-D. All 2,4-D-supplemented media exhibited increased callus induction, suggesting that 2,4-D is an effective growth regulator. The results of this study with modern Egyptian cultivars demonstrated that the response to tissue culture is greatly influenced by the genotype, the type of nutrient medium, and the interaction between them. The most effective explant source for callus indication and plant regeneration is immature embryos.