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Dendrocalamus sericeus is one of bamboo plant species which is very useful for many purposes and the demand of bamboo products has been increasing. It is necessary to rapidly proliferate shoots in vitro and to determine the antioxidant contents application in the medicinal and cosmetic industries. Therefore, the objectives of this study were to investigate the suitable medium for shoot multiplication and to determine antioxidant contents of in vitro shoots. Single-node explants were cultured on Murashige and Skoog (MS) medium supplemented with 0, 1, 2 and 3 mg/L 6-benzyladenine (BA) in combination with 0 and 0.5 mg/L naphthaleneacetic acid (NAA). Then, in vitro regenerated shoot clusters of 2 – 3 shoots were cultured in liquid and on solid MS medium supplemented with 2 mg/l BA in combination with 0, 8 and 15% (v/v) coconut water for four weeks in order to proliferate shoots. The highest shoot formation of 4.67 ± 0.02 and shoot length of 2.51 ± 0.35 cm were observed in liquid MS medium supplemented with 2 mg/l BA in combination with 8% (v/v) coconut water. The antioxidant contents and antioxidant activity of two-week-old in vitro regenerated shoots were determined comparing to field-grown shoots and leaves. The results showed that the higher total phenolic (63.80 ± 1.31 mg GAE/g dry extract) and flavonoid contents (32.18 ± 1.95 mg CE/g dry extract) than field-grown shoots and leaves. In addition, greater DPPH radical scavenging activity (EC50 of 169.17 ± 11.46 µg/ml) and ABTS radical cation decolorization activity (EC50 of 299.39 ± 68.93 µg/ml) were also observed in two-week-old in vitro shoots.