This trial was conducted to assess the impact of medium-chain α-monoglycerides, glycerol monolaurate (GML) supplementation on the growth performance, apparent ileal digestibility coefficient (AID%) of amino acids, intestinal histomorphology, and blood biochemical parameters of broiler chickens. Three-day-old chicks (76.82 g ± 0.40, n = 200) were haphazardly allocated to four experimental groups with five replicates for each (10 chicks/replicate). The treatments consisted of basal diets supplemented with four glycerol monolaurate levels; 0, 1, 3, or 5 g kg−1 (GML0, GML1, GML3, and GML5, respectively). Growth performance was determined at three periods (starter, grower, and finisher). Dietary GML had no significant effect on the growth performance parameters (body weight, weight gain, and feed conversion ratio) through all the experimental periods. GML1 diet increased the AID% of leucine and decreased the AID% of arginine. GML1 diet increased the duodenal and jejunal villous height and the jejunal muscle thickness. GML3 and GML5 diets increased the goblet cell count in the duodenum. GML supplementation increased the serum level of high density lipoprotein (HDL)-cholesterol. GML5 diet increased the serum levels of IgM and interleukin 10 compared to the control group. We could conclude that dietary supplementation of glycerol monolaurate can supplement broiler chicken diets up to 5 g kg−1 to enhance the immune status and intestinal histomorphology of birds with no improving effect on growth performance.