Potato (Solanum tuberosum L.) is a vital crop in Egypt, with the country importing approximately 120,000 MT of seed potatoes annually and exporting over 759,200 MT of ware potatoes. This positions Egypt as Africa's leading potato producer. This study investigated the micropropagation of five potato cultivars (Hermes, Spunta, Lady Rosetta, Cara, and Silana) using in vitro techniques. Nodes from plants were cultured on Murashige and Skoog (MS) medium supplemented with varying concentrations of kinetin (KIN) and Naphthalene Acetic Acid (NAA). Results revealed significant differences among cultivars in survival rates, shoots multiplication, and rooting responses. Hermes explants showed the highest survival rate (100%), with lower but statistically similar survival (96.6%) observed for Silana and Hermes on specific treatments. The highest shoot number (13.6 per jar) was recorded for Hermes cultured on 0.25 mg/L KIN, followed by Hermes and Spunta on 0.5 mg/L NAA. Maximum shoot lengths (7.044 cm and 6.908 cm) were observed with Spunta and Hermes on combined KIN-NAA treatments. Hermes on 0.25–0.5 mg/L KIN exhibited the highest leaf numbers, while root number peaked for Cara and Hermes on 0.5 mg/L NAA + 0.1 mg/L KIN. Spunta exhibited the longest roots (9.667 cm) on 1.0 mg/L NAA + 0.5 mg/L KIN. Acclimatization showed high survival rates, with Lady Rosetta and Spunta achieving 96%. Lady Rosetta also produced the highest minituber weight (40.2 g), while Hermes achieved the highest tuber count (10.7 per plant). These findings emphasize cultivar-specific optimization of growth regulators for large-scale, disease-free potato production in Egypt.