Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, Girona, Girona, Spain.
Mol Reprod Dev. 2013 Nov;80(11):924-35. doi: 10.1002/mrd.22229. Epub 2013 Sep 10.
In vitro culture conditions and certain events during the earliest stages of development are linked to embryonic survival, possibly in a sex-related manner. In vitro-produced (IVP) porcine embryos cultured with glucose (IVC-Glu) or pyruvate-lactate (IVC-PL) were tested for any relationship between the timing of the first embryonic cleavage and development and sex ratio. The embryos were assigned to IVC-Glu or IVC-PL groups and classified depending on the timing of their first cleavage: 24, 26, 30, and 48 hr post-insemination (hpi). They were cultured separately in vitro and evaluated for cleavage rate and pattern, blastocyst rate and stage, cell number, apoptosis, and sex ratio. Regardless of energy source, the percentage of two-cell stage and fragmented embryos at the time of their first cleavage was, respectively, higher and lower in early-cleaving embryos. Those embryos cleaved by 24 hpi developed to blastocysts at a higher rate (IVC-Glu: 37.90 ± 3.06%; IVC-PL: 38.73 ± 4.08%) than those cleaved between 30 and 48 hpi (IVC-Glu: 5.87 ± 3.02%; IVC-PL: 8.41 ± 3.50%). Furthermore, a shift toward males was seen among embryos first cleaved before 30 hpi, versus towards females among those cleaved later. The early-cleaving embryos, only from the IVC-PL group, had a higher proportion of expanded blastocysts (81.05 ± 6.54% vs. 13.33 ± 13.33%) with higher cell numbers than their late-cleaving counterparts. Moreover, a shift toward males only appeared at the blastocyst stage in IVC-PL embryos. These findings confirm that the timing of the first cleavage influences development of IVP porcine embryos in a sex-related manner, and it depends on the main energy source of the in vitro culture medium.
体外培养条件和胚胎发育早期的某些事件与胚胎存活有关,可能与性别有关。用葡萄糖(IVC-Glu)或丙酮酸盐-乳酸盐(IVC-PL)培养的体外生产(IVP)猪胚胎,检测第一次胚胎分裂的时间与性别比例之间的任何关系。胚胎被分配到 IVC-Glu 或 IVC-PL 组,并根据第一次分裂的时间进行分类:受精后 24、26、30 和 48 小时(hpi)。它们在体外分别培养,并评估分裂率和模式、囊胚率和阶段、细胞数量、凋亡和性别比例。无论能量来源如何,第一次分裂时处于两细胞阶段和碎片胚胎的百分比在早期分裂胚胎中分别较高和较低。那些在 24 hpi 时分裂的胚胎以较高的速率发育为囊胚(IVC-Glu:37.90±3.06%;IVC-PL:38.73±4.08%),而那些在 30 至 48 hpi 之间分裂的胚胎则较低(IVC-Glu:5.87±3.02%;IVC-PL:8.41±3.50%)。此外,在 30 hpi 之前第一次分裂的胚胎中出现了向雄性的转变,而在之后分裂的胚胎中则出现了向雌性的转变。只有来自 IVC-PL 组的早期分裂胚胎具有更高比例的扩张囊胚(81.05±6.54%对 13.33±13.33%)和更高的细胞数量,与晚期分裂胚胎相比。此外,只有在 IVC-PL 胚胎中,雄性的转变只出现在囊胚阶段。这些发现证实,第一次分裂的时间以一种与性别相关的方式影响 IVP 猪胚胎的发育,并且它取决于体外培养基的主要能量来源。