Castillo-Martín M, Yeste M, Morató R, Mogas T, Bonet S
Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, Campus Montilivi, E-17071 Girona, Spain.
Reprod Fertil Dev. 2013;25(5):737-45. doi: 10.1071/RD12117.
The objective of the present study was to determine the effects of replacing glucose with pyruvate and lactate during the first 48 h of in vitro culture (IVC) in NCSU-23 medium on embryo development, embryo quality and survival of porcine blastocysts after vitrification. To this end, in vitro-produced (IVP) porcine oocytes were cultured with either glucose for 6 days (IVC-Glu) or pyruvate-lactate from Day 0 to Day 2 and then with glucose until Day 6 (IVC-PyrLac). Blastocysts were vitrified on Day 6 using the Cryotop device and, after warming, survival rate and the apoptosis index were evaluated after 24 h incubation in NCSU-23 medium. No significant differences were observed between IVC-Glu and IVC-PyrLac in terms of cleavage rate, blastocyst yield, total number of cells per blastocyst or the apoptosis index (1.82±0.75% vs 3.18±0.88%, respectively) of non-vitrified embryos. However, a significant increase was seen in hatching/hatched blastocysts in the IVC-PyrLac compared with IVC-Glu treatment group (12.71±1.20% vs 3.54±0.47%, respectively). Regardless of treatment, vitrification impaired the survival rate and the apoptosis index. When comparing both treatments after warming, the percentage of apoptotic cells was significantly higher for blastocysts in the IVC-PyrLac compared with IVC-Glu group (18.55±3.49% vs 9.12±2.17%, respectively). In conclusion, under the conditions of the present study, replacement of glucose with pyruvate-lactate during the first 48 h of culture resulted in a lower cryotolerance of IVP porcine embryos.
本研究的目的是确定在NCSU - 23培养基中体外培养（IVC）的前48小时用丙酮酸和乳酸替代葡萄糖对猪胚胎发育、胚胎质量以及玻璃化后猪囊胚存活率的影响。为此，将体外生产（IVP）的猪卵母细胞分别用葡萄糖培养6天（IVC - Glu），或从第0天至第2天用丙酮酸 - 乳酸培养，然后从第2天至第6天用葡萄糖培养（IVC - PyrLac）。在第6天使用Cryotop装置对囊胚进行玻璃化处理，解冻后，在NCSU - 23培养基中孵育24小时后评估存活率和凋亡指数。在未玻璃化胚胎的卵裂率、囊胚产量、每个囊胚的细胞总数或凋亡指数方面（分别为1.82±0.75%和3.18±0.88%），IVC - Glu和IVC - PyrLac之间未观察到显著差异。然而，与IVC - Glu处理组相比，IVC - PyrLac组孵化/已孵化囊胚显著增加（分别为12.71±1.20%和3.54±0.47%）。无论处理如何，玻璃化都会损害存活率和凋亡指数。解冻后比较两种处理时，IVC - PyrLac组囊胚的凋亡细胞百分比显著高于IVC - Glu组（分别为18.55±3.49%和9.12±2.17%）。总之，在本研究条件下，培养的前48小时用丙酮酸 - 乳酸替代葡萄糖会导致IVP猪胚胎的抗冻性降低。
