To conduct crewed simulation experiments of bioregenerative life support systems on the ground is a critical step for human life support in deep-space exploration. An artificial closed ecosystem named Lunar Palace 1 was built through integrating efficient higher plant cultivation, animal protein production, urine nitrogen recycling, and bioconversion of solid waste. Subsequently, a 105-day, multicrew, closed integrative bioregenerative life support systems experiment in Lunar Palace 1 was carried out from February through May 2014. The results show that environmental conditions as well as the gas balance between O ₂ and CO ₂ in the system were well maintained during the 105-day experiment. A total of 21 plant species in this system kept a harmonious coexistent relationship, and 20.5% nitrogen recovery from urine, 41% solid waste degradation, and a small amount of insect in situ production were achieved. During the 105-day experiment, oxygen and water were recycled, and 55% of the food was regenerated. Key Words: Bioregenerative life support systems (BLSS)—Space agriculture—Space life support—Waste recycle—Water recycle. Astrobiology 16, 925–936.