Female broodstock of Pacific oyster (Crassostrea gigas) were conditioned using different diets to determine the effect of diet on oocyte cryopreservation success. Oocyte biochemical composition, fatty acid profile, and fertilization success before and after cryopreservation were determined. Broodstock were conditioned in 2 stages: a cold preconditioning stage during which animals were either starved or fed live microalgae, and a warm conditioning stage during which animals were again either starved or fed live microalgae, AlgaMac-2000, or a mixture of corn and rice flour. Overall, oysters that were fed microalgae during cold preconditioning produced oocytes with significantly more lipid than oysters that were starved. Similarly, those that were fed during warm conditioning produced oocytes with more lipid than starved oysters, regardless of diet. There was no effect of conditioning regime on the level of polyunsaturated fatty acids (PUFAs) in the polar lipid fraction. However, oysters that were starved or fed AlgaMac-2000 during warm conditioning had higher PUFA levels than oysters fed microalgae or flour. Broodstock conditioning regime had no effect on the fertilization success of oocytes prior to cryopreservation, but those that were fed during cold preconditioning produced oocytes that had significantly higher fertilization success after thawing.