Direct shoot regeneration of two flax cultivars; Blanka (a fiber cultivar) and Giza 8 (a dual purpose cultivar) was conducted in vitro using different explants from the seedling (hypocotyl, cotyledon and root). Factors affecting as well as stages and capacity of direct shoot regeneration and anatomical features of in vitro induced shoots comparing with in vivo shoots were studied. The most remarkable outcomes proved that the used culture medium is the key factor that affecting the shoot regeneration when combined with any other factor (explant type, explant age and genotype). Hypocotyl explant proved to elevate shoot regeneration percentages that varied between 33 to 95%. Three days aged explant bear out the highest percentage of shoot regeneration as compared with explant aged 7 days. The most medium was MS basel medium supplemented with 0.25 mg/l NAA + 0.50 mg/l BA. Inductive flax cv. Blanka showed a higher shoot regeneration percentage as compared with the other genotype flax cv. Giza8. Bud formation was initiated on the surface of the flax seedling hypcotyl as small projection, this projection showed a heavy wrinkled and waved sculpture, later it developed into normal shoot bud with mersitematic dome. The anatomical features of in vivo and in vitro stems and leaves were inversely distinctive as the in vitro stems were wider and maintained thicker epidermal cells as well as thicker cortex and pith whereas having lesser amount of supportive and vascular tissues. Comparing with in vivo leaves, the in vitro leaves were thiner with poor differentiated epidermal, mesophyll and vascular tissues. The in vitro leaves showed bigger and denser stomata area in both leaf surfaces.