The study included the establishment of cell suspension culture of Nigella sativa L. by the presence of the plant growth regulators (2,4-D and Kin) and sulphanilamide. Results showed that the best enhancement media to initiate cell suspension cultures were media containing 2,4-D and Kin at 10-5 , l0-9 molar alternatively which gave the highest rate 95.74 % of callus initiation compared with 98.8 % obtained from the standard media 10-6 molar 2,4-D. the results appeared to approve that the usage of sulphanilamide has the enhancement influence to form the suspension cultures of Nigella sativa when it used with 2,4-D and Kin.
The rate of the callus initiation reached 93.1% in the media containing 75 micromolar sulphanilamide with 10-5, 10-9 molar for 2,4-D and Kin alternatively, it was noticed a good growth in the media containing 100 micromolar of sulphanilamide with 10-5, 10-7 molar for each 2,4-D and Kin, the callus initiation reached 92.9% .
In order to complete achieving the study to find out the contents of cell cultures from the effecting compound (Thymol) comparing them by the thymol extract (control sample). The finding of the cell suspension cultures by using thin layer chromatography (TLC) proved that cell suspension was about to be equal and in some other times it surpass its contents in the comparison compound (Thymol) of the control treatment.Generally, it is clear that the existence of (sulphanilamide) in the nutrient media had given the best results when tested by the (TLC) for the thymol content in cell suspension cultures.