Orostachys cartilaginous A. Bor. has various pharmacological effects, which is mainly distributed in Changbai Mountain area of China, however, the resource of O. cartilaginous is difficult to satisfy the market demand. Plant cell culture can be an alternative approach for alleviating the resource shortage but a problem that relative low yield of bioactive compounds in cell cultures should be solved. Therefore, this study used salicylic acid (SA) as an elicitor to treat 25-day-old bioreactor cultured O. cartilaginous cells and the effects of concentrations and durations of SA treatment on bioactive compound accumulation were investigated to select a suitable strategy of SA elicitation. Results showed that SA concentrations from 25 μM to 150 μM obviously enhanced the bioactive compound accumulation without a decrease of cell biomass. At 100 μM SA, the total polysaccharide (218.5 mg/g DW), phenolic (27.2 mg/g DW), and flavonoid (35.4 mg/g DW) contents reached their maximum levels. Bioactive compound accumulation increased with prolonged SA treatment duration from 0 h to 48, reached the maximum at 48 h, and decreased afterwards. During the SA elicitation, the bursts of hydrogen peroxide (H2O2) and nitric oxide (NO) in cells were detected at the early stage, and peaks of H2O2 and NO contents occurred at 4 h and 6 h, respectively. Furthermore, the maximum contents of flavonoid monomers, including quercetin, kaempferide, epicatechin gallate, quercetin-3-O-glucose, and kaempferol-3-rutinoside were determined at 48 h, thereby speculating that H2O2 and NO might the upstream signaling molecules involved in SA-induced flavonoid synthesis in O. cartilaginous cells. In addition, the O. cartilaginous cell extract (OCE) inhibited growth of Bacillus subtilis and Pseudomonas aeruginosa, and the anti-bacterial activity against B. subtilis was significantly higher than that against P. aeruginosa. Meanwhile, tyrosinase activity was inhibited by the OCE, reaching 61.7% of the inhibitory rate at 1.4 mg/mL OCE, which demonstrated that OCE possessed an anti-melanin ability. This study concluded that 25-day-old bioreactor cultured O. cartilaginous cells can be treated with 100 μM SA for 48 h for the maximum production of polysaccharides, phenolics, and flavonoids; the O. cartilaginous cell cultures could be used as a potential material in the production of anti-bacterial and anti-melanin related products.