The objective of the present experiment was to investigate the effect of stocking density (SD) and dietary supplementation of crystalline tryptophan (Trp) on growth performance and intestinal barrier function in broiler chickens raised in a floor pen. The experiment was conducted using a completely randomized design with a 2 x 2 factorial arrangement consisting of 2 different SD and 2 supplemental levels of dietary Trp. A total of 1,140 Ross 308 broiler chickens at 21 d of age were allotted to 1 of 4 treatments with 5 replicates. Low SD (9 birds/m(2)) and high SD (18 birds/m(2)) were achieved by raising different number of birds per identical floor pen (2.0 m x 2.4 m). The basal diet was formulated with no supplemental Trp in diets to meet or exceed nutrient recommendation of the Ross 308 manual. The calculated concentrations of total Trp and digestible Trp in the basal diet were 0.19 and 0.16%, respectively. The other diet was prepared by adding 0.16% crystalline Trp to the basal diet. Diets were fed to birds for 21 d. At the end of the experiment, 2 birds per replicate were euthanized to collect tissue samples for further analyses. Results indicated that there were no interactions between SD and dietary Trp for all measurements. For the main effects, birds raised at a low SD had greater (P < 0.01) body weight gain, feed intake, and feed efficiency than those raised at a high SD. However, supplementation of dietary Trp had no effect on broiler performance. Furthermore, there were no main effects of SD and dietary Trp on intestinal barrier functions. In conclusion, broiler chickens raised in a floor pen with a high SD (18 birds/m(2)) have decreased growth performance with little changes in intestinal barrier functions. Supplementation of dietary Trp at 0.16% has no positive effect on broiler chickens raised in a floor pen with either a low or high SD.