Optimization of wheat tissue culture is essential for any gene transfer process and for generation of cell suspension lines. To optimize tissue culture conditions for four bread wheat lines (C-D-4, C-D-6, C-D-8, and C-D-9), several factorial experiments based on completely randomized design with 3 replications were conducted. Callus induction potential and regeneration of lines under different hormone treatments were evaluated. For callus induction, ML1G1 medium containing three levels of 2, 4-D (2, 2.4 and 3 mg/l) and for regeneration, MS and N6 medium including different levels of NAA, BAP and Kin was used. With respect to the effect of hormones on the different wheat lines, the results showed that the highest callus induction rate was achieved with C-D-9(82%) and the highest rate of callus induction (60%) was recorded in 2.4 mg/l 2, 4-D for all lines. With respect to embryogenic callus production, ML1C2 media containing 250mg/l AgNO3 produced the highest embryogenic callus percentage in C-D-9 and C-D-8 lines. The highest regeneration rate (29.62%) for C-D-9 lines was obtained in N6 (6) medium containing 1 mg/l IAA + 1 mg/l BA.