The present study investigated the effects of sprouted barley (SB) with different cultivation stages on fermentation characteristics and degradation kinetics in the rumen. The SB was cultivated in three different stages as follows: 0, 4, and 8 days. Dried samples from each cultivation stage of SB were incubated in the rumen buffer at 39 °C for 48 h in quadruplicate with three blanks. Dry matter (DM) and neutral detergent soluble carbohydrate concentrations of SB decreased linearly (p = 0.001) by increasing the cultivation stage, while crude protein, neutral detergent fiber (NDF), acid detergent fiber, and hemicellulose concentrations increased linearly (p ≤ 0.001). Total volatile fatty acid and butyrate in the rumen decreased linearly (p ≤ 0.020) by increasing the cultivation stage, while pH and propionate increased linearly (p < 0.001). The total degradation fraction of DM and NDF increased quadratically (p ≤ 0.003). The fraction degradation rate of DM and NDF decreased linearly (p ≤ 0.001) by increasing the cultivation stage, while the lag phase increased linearly (p ≤ 0.010). The present study concluded that cultivated SB at 4 days was recommended for animal feed due to the highest nutrient degradation in the rumen without any adverse effects on fermentation characteristics.