This study was conducted to optimize callus induction, cell suspension culture, and enhance cichoric acid production in Echinacea purpurea. For this purpose, leaf and root explants were grown in ½ MS medium supplemented with different concentrations of plant growth regulators. The highest percentage of callus induction (100%) was obtained with different concentrations of Kin in combination with 2,4-D and or NAA, in both explants. Then interaction slicing method was applied to analyze the effects of different concentrations of Kin in combination with 2,4-D and or NAA on total phenol contents. The maximum amount of total phenol was observed in 0.5 mg/L kin and 1 mg/L 2,4-D, in both leaf and root-derived callus. Data from HPLC–UV analysis showed that cichoric, chlorogenic, and caffeic acids are present in the callus. While cichoric acid content was 2.11 and 4.73 mg/g dry weight (DW) in leaf explant and its derived callus, it was 3.47 and 5.67 mg/g DW in root explant and its derived callus, respectively. Cell suspension culture was established and treated by AgNO3 and AgNPs (0, 2 and 4 mg/L). Overall, AgNPs’ elicitation was more effective in cichoric acid production compared to AgNO3. The highest amount of cichoric acid (9.54 mg/g DW) was found in root cell suspension culture after 48 h of exposure to 2 mg/L AgNPs. But leaf cell suspension culture reached the maximum cichoric acid level (8 mg/g DW) 72 h after treatment with 2 mg/L AgNPs. The current protocol can be used for the production of cichoric acid from callus and cell suspension cultures.