To obtain oil from bee brood, which was dried using a tray drying method, this study used the supercritical CO2 extraction method. Extraction occurred at temperatures between 40–60 °C and low pressures of 180–220 bar for 1.5 h, with a high pressure of 600 bar for 1 h. The study investigated both the yield and chemical properties of the extracted bee brood oils. Supercritical CO2 extraction of tray-dried bee brood at 600 bar pressure demonstrated higher oil extraction efficiency compared to lower pressures (180–220 bar). At temperatures of 40–60 °C, total phenolic compounds increased while total flavonoids decreased. The extracted oil exhibited antioxidant activity, primarily due to quercetin. Despite decreased acid, iodine, and saponification values, peroxide value slightly increased but remained below 12 meqO2/kg of oil. The make-up of the fatty acids changed. At 600 bar, palmitic and oleic acids were the most common, while myristic, linoleic, and docosadienoic acids decreased. At 600 bar, eicosadienoic acid was absent. The defatted bee brood retained significant essential and non-essential amino acids, indicating its potential for further development as a protein source.