Division of Reproductive Sciences, Oregon National Primate Research Center, Beaverton, Oregon, USA
Biol Reprod. 2010 Oct;83(4):525-32. doi: 10.1095/biolreprod.110.084418. Epub 2010 Jun 2.
The stage at which follicle-enclosed cumulus-oocyte complexes achieve developmental competence in primates is unknown. Therefore, studies were designed to characterize the ability of oocytes in small antral follicles present during the menstrual cycle to spontaneously resume meiosis, fertilize, and support early embryo development. Ovaries were removed from adult rhesus monkeys (n = 12) during the early follicular phase (Days 3-4) of spontaneous cycles. Small antral follicles were divided into five groups according to their diameter; group I: <0.5 mm; group II: 0.5-0.99 mm; group III: 1.0-1.49 mm; group IV: 1.5-1.99 mm; and group V: 2.0-2.5 mm. The cumulus-oocyte complex from healthy small antral follicles (devoid of dark oocytes or granulosa cells) were extracted (n = 199) and cultured for 48 h under different conditions: in TALP (tyrode, albumin, lactate, pyruvate) medium alone, SAGE medium alone, or plus gonadotropins. At 48 h, oocyte meiotic status and diameter were measured after treatment of cumulus-oocyte complexes with hyaluronidase. Cumulus-oocyte complexes derived from follicles of 0.5- to 2-mm diameter contain oocytes that typically reinitiate meiosis in the absence or presence of gonadotropins and fertilize via in vitro fertilization or intracytoplasmic sperm injection. Moreover, the inseminated oocytes can reach the morula stage but arrest. Thus, the ability of these oocytes to complete maturation, as monitored from subsequent embryonic development after fertilization, is suboptimal. Further studies on primate IVM of oocytes from SAFs are warranted in order for them to be considered as an additional, novel source of gametes for fertility preservation in cancer patients.
在灵长类动物中,卵泡包裹的卵丘卵母细胞复合体达到发育能力的阶段尚不清楚。因此,本研究旨在研究在月经周期中存在的小窦卵泡中,卵母细胞自发恢复减数分裂、受精和支持早期胚胎发育的能力。从处于自发周期的早期卵泡期(第 3-4 天)的成年恒河猴(n=12)中取出卵巢。根据卵泡直径将小窦卵泡分为五组:I 组:<0.5mm;II 组:0.5-0.99mm;III 组:1.0-1.49mm;IV 组:1.5-1.99mm;V 组:2.0-2.5mm。从无成熟卵母细胞或颗粒细胞的健康小窦卵泡中提取卵丘卵母细胞复合体(n=199),并在不同条件下培养 48 小时:仅在 TALP(Tyr-,白蛋白,乳酸盐,丙酮酸盐)培养基中、仅在 SAGE 培养基中或在添加促性腺激素的情况下培养。在 48 小时时,用透明质酸酶处理卵丘卵母细胞复合物后,测量卵母细胞减数分裂状态和直径。直径为 0.5-2mm 的卵泡来源的卵丘卵母细胞复合体中含有卵母细胞,在没有或存在促性腺激素的情况下,这些卵母细胞通常可以重新启动减数分裂,并通过体外受精或胞浆内精子注射受精。此外,受精的卵母细胞可以达到桑椹胚阶段,但停滞不前。因此,这些卵母细胞从受精后胚胎发育中监测的成熟能力并不理想。需要进一步研究灵长类 SAF 中的卵母细胞 IVM,以便将其作为癌症患者生育力保存的另一种新的配子来源。