This study was conducted in the Plant Cell and Tissue Culture Laboratory of the Department of Life Sciences at the College of Education for Pure Sciences at the University of Diyala during the period from September 2020 to July 2021. This study was carried out with the aim of obtaining typical suspended cell cultures. Of the broccoli plant Brassica Oleracea of medicinal and pharmaceutical value, which is derived from the callus of the subcotyledonous stems of broccoli grown on MS medium supplemented with different concentrations of 2,4-(2,4-D) Dichlorophenoxy acetic acid (interlaced with Benzyl adenine (BA) and then follow-up The results of its cultivation, the results indicate the following. The stems grown on MS medium supplemented with growth regulators have superiority and ability to induce callus compared to their peers in the control treatment. It was found that the MS medium supplemented with concentrations of 1.5,2.00 mg.l-1,4-D + 0.5 mg.l-1 BA stimulated callus induction and achieved the highest value of 100% compared to the control treatment whose induction was 0%, and the percentage of Induction to 50% in the treatment at a concentration of 0.5,1.00 mg.l ¬¬-1,4-D + 0.5 mg.l-1 BA. The results also indicate that the highest average fresh weight of the callus was recorded when treatment of 1.5.2.00 mg.l ¬¬-1 of growth regulators, which reached 1.166,2.343 g.piece -1 in a row, outperforming the treatment with a concentration of 0.5,1.00 mg. L-1 2,4-D + 0.5 mg.L-1 BA, whose value started decreasing to 0.313,0.860 g.pc-1, respectively. The results showed that the concentration exceeded 2.00 mg. 1 liter of 2,4-D + 0.5 of BA added to it nano iron oxide was significantly higher than the rest of the concentrations, as it recorded the highest mean of 4.453 gm of fresh weight compared to the control treatment of 2.343 gm. for the stem and amounted to 4.333 gm compared to the control treatment which amounted to 2.650 gm for the leaf the.