Bishop's seed (Trachyspermum ammi L.) is a vital neutraceutial plant used as spice and also used as medicinal plant for curing different ailments and diseases of humans and animals. The study presents the in vitro regeneration of Bishop's seed using cotyledonary node explant taken from in vitro germinated seeds. All mediums used in this study were comprised of MS (0.44%) medium having 3.0%sucrose and 0.65% agar with pH of 5.8. Explants were inoculated on MS medium enriched with 0.10-1.60 mg/L Thidiazuron (TDZ) alone or in combination with 0.10 mg/L IBA (Indole-3-butyric acid) for seven weeks under 16/8 light photoperiod provided with cool-white fluorescent lamps. Thereafter, explants were sucultured on MS medium without any plant growth regulators (PGRs) for five weeks more under similar culture conditions. Results revealed 100% shoot regeneration frequency with 42.85-100.00%. The shoot count and shoot length ranged 3.43-19.40 and 1.04-2.74 cm respectively. The highest shoot count (19.40) and shoot length (2.74 cm) were observed on MS medium enriched with 0.20 mg/L TDZ and 0.20 mg/L TDZ+0.10 mg/L IBA respectively. Relatively low rooting was recorded on IBA containing medium with low adaptation percentage of plantlets in pots containing peat moss.