The purpose of this study was to investigate the potential benefits of yeast cell wall (YCW) on gastrointestinal development of weaned calves. A total of 20 healthy Holstein male calves (92 ± 8.29 kg of body weight and 60 ± 5 ddays of age) were randomly allocated into 2 groups: CON [no YCW (Angel Yeast Co., Ltd.)], and YCW (accounted for 0.16% of the basal diet). The dietary concentrate to roughage ratio was 40:60, and all calves were fed regularly twice a day at 09:00 and 16:00, besides, calves. Calves had ad libitum access to water. The experiment lasted for 60 ddays. The results showed that in the ventral sac of the rumen, compared to CON, calves fed YCW showed higher lengths (P < 0.05), widths (P < 0.05), and surface areas (P < 0.05) of papillae. For the dorsal sac of the rumen, the muscularis thickness was thicker (P < 0.05) in YCW compared with CON. In ileum, the villus height of YCW calves was higher (P < 0.05) than that in CON. Calves supplemented with YCW also showed a higher (P < 0.05) villus height to crypt depth ratio in the ileum. YCW calves exhibited a greater (P < 0.05) thickness of wall in the duodenum and jejunum. Calves supplemented with YCW improved (P < 0.05) the mRNA expression of Claudin1 in ileum. In addition, calves fed YCW exhibited higher (P < 0.05) mRNA expression of Occludin in the jejunum and ileum. YCW increased (P < 0.05) the contents of secretory immunoglobulin A in jejunum and ileum of calves. In conclusion, the basal diet supplemented with YCW could improve the gastrointestinal development of weaned calves.