Bioactive compounds of plant cell culture extracts are currently being looked at for its potential for the development of nutraceutical products. Tomato callus is known to have cytoprotective activity. The availability for explants and its callus production are two important things. This paper reports the effect of different strengths of medium on seed germination and seedling growth of tomato cv. Permata and sucrose effects on their biomass callus. Seeds were grown on 4 kind of germination media. Hypocotyl were taken as explants for callus induction in MS medium supplemented by 2.0 mg/l BA and 0.2 mg/l NAA, 3% sucrose, and subcultured after 20 days on medium with 6 variations of sucrose. The highest values for tomato hypocotyl’s height were obtained from the ½ and ¼ MS, while the highest fresh weight was obtained from the MS 0. Both the fresh weight and dry weight of the callus increased along with increasing sucrose concentration. The explant age and biomass need to be a concern regarding what kind of callus that will be produced. Application of 5% sucrose concentration is recommended for callus production in this study.