Cryogenic pressure vessel’s low-cycle fatigue performance is greatly affected by cold forming and warm forming. Tensile experiments and low-cycle fatigue experiments of metastable austenitic stainless steel S30408 at liquid-nitrogen temperature (77 K) were conducted to analyze the influences of pre-strain (0 and 0.35) and pre-strain temperature (293 K and 363 K) on the cryogenic low-cycle fatigue performance. Due to 0.35 pre-strain resulting in the increase in dislocation density and formation of strain-induced body centered cubic martensite, 293 K pre-strain specimen shows higher initial cyclic stress amplitude, smaller fatigue striation width and longer low-cycle fatigue life than base metal specimen at 77 K. When the pre-strain temperature changes from 293 K to 363 K, the austenite phase owns higher stability, martensitic transformation is blocked, the initial cyclic stress amplitude decreases, the fatigue striation width increases, and the strengthening effects on the cryogenic low-cycle fatigue life caused by 0.35 pre-strain is weakened. In general, 0.35 pre-strain shows significant strengthening effects on the S30408’s cryogenic low-cycle fatigue performance, and the effects are limited by the pre-strain temperature.