The invention discloses a pig oocyte in-vitro maturation culture solution and a preparation method and application thereof, and belongs to the technical field of oocyte in-vitro maturation culture. The problems of low in-vitro maturation rate and development rate of porcine oocytes currently are solved. The pig oocyte in-vitro maturation culture solution is prepared from a basal culture solution TCM-199, penicillin, streptomycin, NaHCO3, 4-hydroxyethylpiperazine ethane sulfonic acid, polyvinyl alcohol, sodium pyruvate, insulin, cysteine, epidermal growth factor, porcine follicular fluid, pregnant mare serum gonadotropin, human chorionic gonadotropin and tannic acid. According to the pig oocyte in vitro maturation culture solution, the oocyte in-vitro maturation quality, the cumulus cell diffusion index, the parthenogenetic embryo blastocyst rate and blastocyst total cell number, and the in-vitro fertilization embryo cleavage rate, blastocyst rate and blastocyst total cell number can beimproved.