Schramm R D, Bavister B D
Wisconsin Regional Primate Research Center, Madison 53715, USA.
Hum Reprod. 1995 Apr;10(4):887-95. doi: 10.1093/oxfordjournals.humrep.a136056.
Specific aims of this study were to determine effects of granulosa cells and gonadotrophins on the meiotic and developmental competence in vitro of oocytes from non-stimulated rhesus monkeys. Oocytes (368) were obtained from small antral follicles (class 1: 700-999 microns; class 2: 1000-2500 microns) dissected from excised ovaries of 10 follicular and three luteal phase monkeys. Oocytes were cultured in one of four treatment groups: (i) granulosa cells (4 x 10(6) live cells/ml) + gonadotrophins (5 micrograms/ml follicle stimulating hormone and 10 micrograms/ml luteinizing hormone), (ii) granulosa cells alone, (iii) gonadotrophins alone, or (iv) controls (no granulosa cells or gonadotrophins). Mature oocytes were inseminated and cultured until arrest of embryo development. Meiotic and developmental capacity was greater (P < or = 0.05) for oocytes from class 2 compared with class 1 follicles and from luteal compared with follicular phase monkeys. Culture of oocytes with gonadotrophins alone during in-vitro maturation improved (P < or = 0.01) activation and cleavage through the 5-8-cell stage (46.5%). Culture with granulosa cells alone during in-vitro maturation augmented progression of embryos to the morula stage (6.3%). The combination of granulosa cells+gonadotrophins enhanced (P < or = 0.01) nuclear maturation, but had no effects on developmental capacity beyond those of either treatment alone. In conclusion, both granulosa cells and gonadotrophins during in-vitro maturation have stimulatory effects on the meiotic and developmental competence of oocytes from non-stimulated macaques in vitro.
本研究的具体目的是确定颗粒细胞和促性腺激素对未受刺激的恒河猴卵母细胞体外减数分裂和发育能力的影响。从10只处于卵泡期和3只处于黄体期的猴子切除的卵巢中解剖出小卵泡（1类：700 - 999微米；2类：1000 - 2500微米），获得368枚卵母细胞。卵母细胞在四个处理组之一中培养：（i）颗粒细胞（4×10⁶活细胞/毫升）+促性腺激素（5微克/毫升促卵泡激素和10微克/毫升促黄体生成素），（ii）单独的颗粒细胞，（iii）单独的促性腺激素，或（iv）对照组（无颗粒细胞或促性腺激素）。成熟卵母细胞受精并培养至胚胎发育停滞。与1类卵泡的卵母细胞相比，2类卵泡的卵母细胞以及与卵泡期猴子相比，黄体期猴子的卵母细胞的减数分裂和发育能力更强（P≤0.05）。在体外成熟期间单独用促性腺激素培养卵母细胞可改善（P≤0.01）激活和分裂至5 - 8细胞期（46.5%）。在体外成熟期间单独用颗粒细胞培养可增加胚胎发育至桑葚胚期的比例（6.3%）。颗粒细胞 + 促性腺激素的组合增强了（P≤0.01）核成熟，但对发育能力的影响不超过单独任何一种处理。总之，体外成熟期间颗粒细胞和促性腺激素对未受刺激的猕猴卵母细胞的减数分裂和发育能力均有刺激作用。
