The oxidative activities of hydroponically produced Chinese kale (Brassica oleracea var. alboglabra L.) seedlings grown under different concentrations of selenium (Se) biofortification with sodium selenite (Na2SeO3) at 0, 5, 10, 15, and 30 mg·Se/L and different ages (5, 10, and 15 days after germination) were determined by the electron transfer method (DPPH and FTC). Highest antioxidant activities of Se-enriched Chinese kale seedlings (Se-KS) were found in seedlings grown on 30 mg·Se/L, 15 days after germination. The physicochemical contents (moisture, crude fibre, ash, crude protein, and carbohydrate), total Se contents, and Se bioaccessibility and antioxidant activities (after treatment with an in vitro simulated gastrointestinal digestion process) between Se-KS and a control of regular Chinese kale seedlings (R-KS) were compared. The crude protein and total Se contents of the air-dried matter in Se-KS were higher (37 ± 1% and 433 ± 22 mg·Se/kg) than those of the R-KS (33 ± 2% and 17 ± 3 mg·Se/kg). Total Se content and antioxidant activities were strongly correlated in the range from 0.93 to 0.99. Testing acute toxicity, a single dose of the Se-KS was orally administrated to rats at 1250, 2500, and 5000 mg/kg·b.w., which consisted of 0.54, 1.08, and 2.16 mg Se/kg·b.w., respectively. These results demonstrated that Se-KS did not create any toxicological signs and mortality. No abnormal pathological changes in the liver, kidney, and heart were found. The results suggested that the Se-KS could be a good source of organo Se for development as a new Se supplementary product.