Sankai T, Shimizu K, Cho F, Yoshikawa Y
Tsukuba Primate Center for Medical Science, National Institute of Health of Japan, Ibaraki.
Jikken Dobutsu. 1994 Jan;43(1):45-50.
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.
使用4只雌性日本猕猴，通过多次注射孕马血清促性腺激素（PMSG）和单次注射人绒毛膜促性腺激素（hCG）诱导卵泡生长，并从卵泡中收集卵母细胞。注射200国际单位的PMSG六次或八次以刺激卵泡生长，在这两种情况下均充分诱导了卵泡生长。从两只雌性猕猴的94个卵泡中回收了47个（50.0%）卵母细胞，并在10小时内从犬山市运送到筑波市。到达时观察到5个成熟卵母细胞和42个未成熟卵母细胞。42个未成熟卵母细胞中有13个（31.0%）在含胎牛血清（FCS）和PMSG的TCM - 199培养基中培养24小时（包括运输时间）后排出了第一极体。18个成熟卵母细胞用于体外受精（IVF）。将预先孵育2小时的冷冻解冻食蟹猴精子转移到含有日本猕猴卵母细胞的液滴中。基于雄原核和/或第二极体的存在，判断16个卵母细胞（88.9%）成功受精，其中12个卵母细胞随后发生分裂，3个发育到6细胞阶段。因此，我们成功诱导了卵泡生长和卵母细胞成熟，并使用食蟹猴精子和日本猕猴卵母细胞实现了体外受精。
