Lycium Fruit (fruits, “Kukoshi”) and Lycium Bark (root barks, “Jikoppi”) are source of the oriental medicines, and Lycium chinense Miller and L. barbarum are its original plants. Moreover, its leaves and stems also contain many kinds of antioxidative polyphenolic compounds such as chlorogenic acids. There are few reports on the production of secondary metabolites by the tissue culture of L. chinense. Therefore, we found the optimal condition of phytohormones for callus induction from stems of L. chinense. Moreover, we determined the antioxidant compounds contained in calli of L. chinense. The sterilized stems of cultivars of L. chinense were cultured on Murashige and Skoog (MS), Gamborg B5 (B5), or Lloyd and McCown’s Woody Plant basal media (WP) containing the combination of 10 μM naphthylacetic acid (NAA), 2,4-dichlorophenoxyacetic acid(2,4-D) or indolebutyric acid (IBA), plus 10 μM kinetin (KIN), thidiazuron (TDZ) or benzyladenine (BA). Methanol extracts of calli were analyzed by LC-ESI-MS and its antioxidant activity was evaluated by ORAC method. In the results, the callus induction rate in B5 basal medium was higher than that of MS and WP. In all combinations of auxins and cytokinins contained in B5 basal medium, calli were well-induced, and the callus induction rate in the conditions containing NAA were higher than 2,4-D and IBA. Besides, the methanol extracts of the calli induced on WP were high antioxidant activity rather than MS and B5 and contained chlorogenic acid (3-CQA) and kukoamine derivatives. We have newly reported the calli of L. chinense produced some useful antioxidant compounds.