In Vitro Fertilization Using Japanese Monkey (Macaca fuscata) Eggs and Cynomolgus Monkey (Macaca fascicularis) Frozen/Thawed Spermatozoa

Abstract

Using four female Japanese monkeys, follicular growth was induced by multiple PMSG and single hCG injections, and oocytes were collected from their follicles. PMSG, 200 IU, was injected six or eight times to stimulate follicular growth, and in both cases growth of the follicles was adequately induced. Forty-seven (50.0%) oocytes were recovered from 94 follicles of two of the females and transported from Inuyama-shi to Tsukuba-shi in 10 hours. Five mature oocytes and 42 immature oocytes were observed upon arrival. Thirteen (31.0%) of the 42 immature oocytes released the first polar body in TCM-199 medium containing FCS and PMSG after 24 hour-culture, including transport time. Eighteen mature oocytes were used for in vitro fertilization (IVF) . Frozen and thawed cynomolgus monkey spermatozoa preincubated for 2 hours were transferred into the drops containing the Japanese monkey oocytes. Sixteen oocytes (88.9%) were judged to have been successfully fertilized based on the presence of a male pronucleus and/or second polar body, and 12 oocytes then cleaved, and 3 developed into the 6-cell stage. Thus, we succeeded in inducing follicular growth and oocyte maturation, and in achieving IVF using cynomolgus monkey spermatozoa and Japanese monkey oocytes.