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Abstract：
BACKGROUND: The common marmoset, Callithrix jacchus, is an invaluable model in biomedical research. Its use includes genetic engineering applications, which require manipulations of oocytes and production of embryos in vitro. To maximize the recovery of oocytes suitable for embryo production and to fulfil the requirements of the 3R principles to the highest degree possible, optimization of ovarian stimulation protocols is crucial. Here, we compared the efficacy of two hormonal ovarian stimulation approaches: 1) stimulation of follicular growth with hFSH followed by triggering of oocyte maturation with hCG (FSH + hCG) and 2) stimulation with hFSH only (FSH-priming).
METHODS: In total, 14 female marmosets were used as oocyte donors in this study. Each animal underwent up to four surgical interventions, with the first three performed as ovum pick-up (OPU) procedures and the last one being an ovariohysterectomy (OvH). In total, 20 experiments were carried out with FSH + hCG stimulation and 18 with FSH-priming. Efficacy of each stimulation protocol was assessed through in vitro maturation (IVM), in vitro fertilization (IVF) and embryo production rates.
RESULTS: Each study group consisted of two subgroups: the in vivo matured oocytes and the oocytes that underwent IVM. Surprisingly, in the absence of hCG triggering some of the oocytes recovered were at the MII stage, moreover, their number was not significantly lower compared to FSH + hCG stimulation (2.8 vs. 3.9, respectively (ns)). While the IVM and IVF rates did not differ between the two stimulation groups, the IVF rates of in vivo matured oocytes were significantly lower compared to in vitro matured ones in both FSH-priming and FSH + hCG groups. In total, 1.7 eight-cell embryos/experiment (OPU) and 2.1 eight-cell embryos/experiment (OvH) were obtained after FSH + hCG stimulation vs. 1.8 eight-cell embryos/experiment (OPU) and 5.0 eight-cell embryos/experiment (OvH) following FSH-priming. These numbers include embryos obtained from both in vivo and in vitro matured oocytes.
CONCLUSIONS: A significantly lower developmental competence of the in vivo matured oocytes renders triggering of the in vivo maturation with hCG as a part of the currently used FSH-stimulation protocol unnecessary. In actual numbers, between 1 and 7 blastocysts were obtained following each FSH-priming. In the absence of further studies, FSH-priming appears superior to FSH + hCG stimulation in the common marmoset under current experimental settings.
METHODS: In total, 14 female marmosets were used as oocyte donors in this study. Each animal underwent up to four surgical interventions, with the first three performed as ovum pick-up (OPU) procedures and the last one being an ovariohysterectomy (OvH). In total, 20 experiments were carried out with FSH + hCG stimulation and 18 with FSH-priming. Efficacy of each stimulation protocol was assessed through in vitro maturation (IVM), in vitro fertilization (IVF) and embryo production rates.
RESULTS: Each study group consisted of two subgroups: the in vivo matured oocytes and the oocytes that underwent IVM. Surprisingly, in the absence of hCG triggering some of the oocytes recovered were at the MII stage, moreover, their number was not significantly lower compared to FSH + hCG stimulation (2.8 vs. 3.9, respectively (ns)). While the IVM and IVF rates did not differ between the two stimulation groups, the IVF rates of in vivo matured oocytes were significantly lower compared to in vitro matured ones in both FSH-priming and FSH + hCG groups. In total, 1.7 eight-cell embryos/experiment (OPU) and 2.1 eight-cell embryos/experiment (OvH) were obtained after FSH + hCG stimulation vs. 1.8 eight-cell embryos/experiment (OPU) and 5.0 eight-cell embryos/experiment (OvH) following FSH-priming. These numbers include embryos obtained from both in vivo and in vitro matured oocytes.
CONCLUSIONS: A significantly lower developmental competence of the in vivo matured oocytes renders triggering of the in vivo maturation with hCG as a part of the currently used FSH-stimulation protocol unnecessary. In actual numbers, between 1 and 7 blastocysts were obtained following each FSH-priming. In the absence of further studies, FSH-priming appears superior to FSH + hCG stimulation in the common marmoset under current experimental settings.
摘要：
背景：普通的马莫西，Callithrixjacchus,是生物医学研究中的宝贵模型。它的用途包括基因工程应用，这需要在体外操作卵母细胞和生产胚胎。为了最大限度地回收适合胚胎生产的卵母细胞，并尽可能最大程度地满足3R原则的要求，优化卵巢刺激方案至关重要。这里,我们比较了两种激素卵巢刺激方法的疗效:1)用hFSH刺激卵泡生长,然后用hCG(FSH+hCG)触发卵母细胞成熟,2)用hFSH刺激(FSH-priming).
方法：总共，在这项研究中，将14只雌性猕猴用作卵母细胞供体。每只动物都经历了多达四次手术干预，前三个作为卵子拾取（OPU）程序和最后一个是卵巢子宫切除术（OvH）。总的来说,用FSH+hCG刺激进行20个实验,用FSH引发进行18个实验。通过体外成熟（IVM）评估每种刺激方案的功效，体外受精（IVF）和胚胎生产率。
结果：每个研究组由两个亚组组成：体内成熟卵母细胞和接受IVM的卵母细胞。令人惊讶的是,在没有hCG触发的情况下，一些回收的卵母细胞处于MII期，此外,与FSH+hCG刺激相比，它们的数量并没有显著降低(2.8vs.3.9，分别为（ns））。虽然两个刺激组之间的IVM和IVF率没有差异，在FSH引发组和FSH+hCG组中,体内成熟卵母细胞的IVF率显著低于体外成熟卵母细胞.总的来说,1.7八细胞胚胎/实验（OPU）和2.1八细胞胚胎/实验（OvH）后获得FSH+hCG刺激与1.8FSH引发后的八细胞胚胎/实验（OPU）和5.0八细胞胚胎/实验（OvH）。这些数字包括从体内和体外成熟卵母细胞获得的胚胎。
结论：体内成熟卵母细胞的发育能力显著降低，使得用hCG作为目前使用的FSH刺激方案的一部分来触发体内成熟。在实际数字中，每次FSH引发后获得1至7个胚泡。在没有进一步研究的情况下，在当前的实验设置下，在普通mar猴中，FSH引发似乎优于FSHhCG刺激。
方法：总共，在这项研究中，将14只雌性猕猴用作卵母细胞供体。每只动物都经历了多达四次手术干预，前三个作为卵子拾取（OPU）程序和最后一个是卵巢子宫切除术（OvH）。总的来说,用FSH+hCG刺激进行20个实验,用FSH引发进行18个实验。通过体外成熟（IVM）评估每种刺激方案的功效，体外受精（IVF）和胚胎生产率。
结果：每个研究组由两个亚组组成：体内成熟卵母细胞和接受IVM的卵母细胞。令人惊讶的是,在没有hCG触发的情况下，一些回收的卵母细胞处于MII期，此外,与FSH+hCG刺激相比，它们的数量并没有显著降低(2.8vs.3.9，分别为（ns））。虽然两个刺激组之间的IVM和IVF率没有差异，在FSH引发组和FSH+hCG组中,体内成熟卵母细胞的IVF率显著低于体外成熟卵母细胞.总的来说,1.7八细胞胚胎/实验（OPU）和2.1八细胞胚胎/实验（OvH）后获得FSH+hCG刺激与1.8FSH引发后的八细胞胚胎/实验（OPU）和5.0八细胞胚胎/实验（OvH）。这些数字包括从体内和体外成熟卵母细胞获得的胚胎。
结论：体内成熟卵母细胞的发育能力显著降低，使得用hCG作为目前使用的FSH刺激方案的一部分来触发体内成熟。在实际数字中，每次FSH引发后获得1至7个胚泡。在没有进一步研究的情况下，在当前的实验设置下，在普通mar猴中，FSH引发似乎优于FSHhCG刺激。
