Bioaerosol concentrations and sizes in indoor environment are associated with human health. Among various indoor environments, the knowledge of the bioaerosol characteristics in underground garages remains unclear. In this study, both DAPI (4′,6-diamidino-2-phenylindole) staining and culturable methods were performed to determine the concentrations and size distributions of bioaerosols in two typical underground garages (an office building (UBOB) and a commercial building (UBCB)) in Xi'an, China. Results indicated that bioaerosol concentrations showed seasonal variations in two garages. The total bioaerosol concentrations were higher in summer (UBOB: (1.07 ± 0.29) ✕105 cells/m3; UBCB: (0.59 ± 0.23) ✕105 cells/m3) than winter (UBOB: (0.34 ± 0.17) ✕105 cells/m3; UGCB: (0.38 ± 0.15)✕105 cells/m3). Higher concentrations of bioaerosols were also found during the rush hour than other times due to the elevated traffic volumes. The spatial distribution of bioaerosols was relatively even for most sampling sites in two garages. Moreover, over 80% of airborne culturable fungi and over 65% of airborne culturable bacteria were in respirable size range (<4.7 μm). Importantly, airborne culturable fungal concentrations in two garages exceeded the proposed thresholds by the World Health Organization (WHO) (500 CFU/m3), with the exceeding standard rate (ESR) of 80.28% and 82.60%, respectively, at UGOB and UGCB, implying that necessary control measures should be taken to reduce microorganism contamination in the air of underground garages. The present results may provide the scientific basis for better understanding pollution characteristics of bioaerosols and formulating prevention and control measures in underground garages.