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体外生产的猪胚胎的胚胎发育和性别比例受添加到培养基中的能量底物和透明质酸的影响。

Embryo development and sex ratio of in vitro-produced porcine embryos are affected by the energy substrate and hyaluronic acid added to the culture medium.

作者信息

Torner Eva, Bussalleu Eva, Briz M Dolors, Yeste Marc, Bonet Sergi

机构信息

Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, Campus Montilivi, s/n, 17071 Girona, Spain.

Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, University Campus, 08193 Bellaterra, Barcelona, Spain.

出版信息

Reprod Fertil Dev. 2014;26(4):570-7. doi: 10.1071/RD13004.

Abstract

In the present study, the effects of replacing glucose with pyruvate-lactate and supplementing these in vitro culture (IVC) media with hyaluronic acid (HA) on porcine embryo development and sex ratio were examined. The in vitro-produced (IVP) porcine embryos were cultured in NCSU-23 medium with 0.0, 0.5 or 1.0mgmL(-1) HA, and with either 5.55mM glucose (IVC-Glu) or pyruvate (0.17mM)-lactate (2.73mM) from 0 to 48h post insemination (h.p.i.) and then with glucose from 48 to 168h.p.i. (IVC-PL). Those embryos cultured with IVC-PL had significantly higher blastocyst rates (23.7±1.5%) than those cultured with IVC-Glu (14.27±2.75%). At 1.0mgmL(-1), HA tended to skew the sex ratio of blastocysts towards males in those embryos cultured in IVC-PL, and led to a significant decrease in the blastocyst rate compared with embryos cultured in the presence of 0.5 and 0.0mgmL(-1) HA and IVC-Glu (4.28±0.28% vs 11.01±1.42% and 10.14±2.77%, respectively) and IVC-PL (14.37±1.35% vs 20.96±2.85% and 22.99±1.39%, respectively). In contrast, there were no significant differences in the total cell number per blastocyst or in apoptosis rates. In conclusion, pyruvate and lactate were the preferred energy substrates in the early stages of IVP porcine embryos. Moreover, 1.0mgmL(-1) HA significantly decreased the percentage of blastocyst rates in both the IVC-Glu and IVC-PL groups, but only by a preferential loss of female embryos for those cultured in IVC-PL.

摘要

在本研究中,检测了用丙酮酸-乳酸替代葡萄糖以及在这些体外培养(IVC)培养基中添加透明质酸(HA)对猪胚胎发育和性别比例的影响。体外生产(IVP)的猪胚胎在含有0.0、0.5或1.0mg/mL HA的NCSU-23培养基中培养,在授精后0至48小时(h.p.i.)添加5.55mM葡萄糖(IVC-Glu)或丙酮酸(0.17mM)-乳酸(2.73mM),然后在48至168h.p.i.添加葡萄糖(IVC-PL)。与用IVC-Glu培养的胚胎(14.27±2.75%)相比,用IVC-PL培养的胚胎囊胚率显著更高(23.7±1.5%)。在1.0mg/mL时,HA倾向于使IVC-PL培养的胚胎中囊胚的性别比例偏向雄性,并且与在含有0.5和0.0mg/mL HA以及IVC-Glu(分别为4.28±0.28%对11.01±1.42%和10.14±2.77%)和IVC-PL(分别为14.37±1.35%对20.96±2.85%和22.99±1.39%)培养的胚胎相比,导致囊胚率显著降低。相比之下,每个囊胚的总细胞数或凋亡率没有显著差异。总之,丙酮酸和乳酸是IVP猪胚胎早期阶段的首选能量底物。此外,1.0mg/mL HA显著降低了IVC-Glu和IVC-PL组的囊胚率百分比,但仅在IVC-PL培养的胚胎中通过雌性胚胎的优先损失。

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