Digitalis lanata is commonly used as a drug for congestive heart diseases. The goal of this work is to develop bioactive components from D. lanata cell suspension culture. To achieve this purpose, we used progesterone as a precursor, KCl, and mannitol as elicitors, as a significant biotechnological tool to produce secondary metabolites from cell culture system. The results demonstrated that the medium containing 1.0 mg/l NAA + 1.0 mg/l BAP was more suitable for forming friable calli with acceptable texture for cell suspension culture. The medium B, which contained 1.0 mg/l NAA + 0.5 mg/l BAP, produced higher callus fresh weight and viable cell number/ml than medium A, and the greatest cell growth occurred between days 12 and 15. The linearity and recovery of digitoxin and digoxin in HPLC were within acceptable limits. The addition of 40 mM of KCl resulted in the highest digitoxin value of 5.632 mg/g DW compared to the control of 0.038 mg/g DW. In the case of digoxin, the cell suspension control without any addition showed a maximum value of 671 g/g DW.