Culture of Arthrospira platensis (Spirulina platensis) in human urine was investigated to get valuable biomass. NO3-N was the proper N source, in comparison with other N source, including urea, NH4-N and NO2-N. As a result, aerobic nitrification of human urine was performed, with above 93.6% nitrification percentage finally achieved with total-N (TN) load of 46.52 mg/(L·d), in which Arthrospira platensis was successfully grown. The main compositions of the obtained biomass are close to those in Zarrouk medium. Thus, it is possible to culture Arthrospira platensis in nitrified human urine for food production within bioregenerative life support systems (BLSSs).