The effects of broiler breeder age (26, 28, and 30 wk) and incubator relative humidity (43, 53, and 63% RH) on yolk sac weight and composition in broiler hatching eggs between 16 and 19 d of incubation were evaluated. Percentage yolk sac weight was reduced in eg-gs from the youngest (26 wk) breeder hens and in eggs incubated at 53% RH. Percentage yolk moisture, in eggs from the youngest hens, was lowest at 16 d and was highest at 19 d, and percentage yolk lipid was highest at 16 and 17 d of incubation in eggs from 30-wk-old hens. Percentage yolk lipid content was higher at 63% RH compared with 53% RH in eggs from 26-wk-old breeders but was lower in eggs incubated at a 43% RH compared with 53 and 63% RH from 30-wk-old breeders. Yolk stearic and arachidonic acid concentrations were higher and myristic acid concentration was lower in eggs from the 26-wk-old hens. Conversely, the highest yolk concentrations of linolenic and palmitoleic acids occurred in eggs from 30- and 28-wk-old breeders, respectively. At 16, 17, 18, and 19 d of incubation, yolk oleic acid concentrations were significantly, but variably, affected by breeder age, whereas linoleic acid concentration was highest in 26-wk-old breeders only at 19 d. Palmitic acid concentration at 17 d was lower in eggs incubated at 53% RH compared with 43 and 63% RH. Rate of yolk uptake from the yolk sac by embryos may be increased by incubating eggs at 53% RH, and the effects of changes in RH between 43 and 63% on yolk lipid content are influenced by breeder age.