Donnay I, Feugang J M, Bernard S, Marchandise J, Pampfer S, Moens A, Dessy F
Veterinary Unit, Catholic University of Louvain, Louvain-la-Neuve, Belgium.
Zygote. 2002 Aug;10(3):189-99. doi: 10.1017/s0967199402002253.
Although toxic for early stages of embryo development, glucose is a physiological metabolic substrate at the morula and blastocyst stages. We evaluated the effect of adding 5.5 mM glucose from the morula stage on bovine blastocyst development and quality. In vitro matured and fertilised bovine oocytes were cultured in modified Synthetic Oviduct Fluid medium containing 5% fetal calf serum, but without added glucose, up to day 5 post-insemination (pi). Morulae were selected and further cultured in the presence or absence of 5.5 mM glucose. Blastocyst and hatched blastocyst rates were recorded. Oxygen, glucose and pyruvate uptakes as well as lactate release were evaluated. The quality of the resulting blastocysts was evaluated by the cell allocation to the inner cell mass (ICM) and trophectoderm (TE) and by the apoptotic index. Adding glucose increased the blastocyst rate at day 8 pi (80% vs 65%) but had no impact on hatching rate (25% vs 28%). A 22% decrease in oxygen uptake was observed in the presence of glucose, concomitant with an increase in lactate release, although no change was observed in pyruvate uptake. A slight decrease in blastocyst cell number was observed at day 7 in the presence of glucose while neither the ICM/TE cell ratio nor the apoptotic index were affected. In conclusion, adding 5.5 mM glucose from the morula stage has a limited impact on blastocyst rate and quality although important modifications were observed in embryo metabolism. It remains to be determined whether those modifications could influence embryo viability after transfer.
尽管葡萄糖对胚胎发育的早期阶段有毒性,但在桑椹胚和囊胚阶段它是一种生理性代谢底物。我们评估了从桑椹胚阶段添加5.5 mM葡萄糖对牛囊胚发育和质量的影响。将体外成熟和受精的牛卵母细胞在含有5%胎牛血清但未添加葡萄糖的改良合成输卵管液培养基中培养至授精后第5天(pi)。选择桑椹胚并在有或无5.5 mM葡萄糖的情况下进一步培养。记录囊胚和孵化囊胚率。评估氧气、葡萄糖和丙酮酸摄取以及乳酸释放。通过分配到内细胞团(ICM)和滋养外胚层(TE)的细胞以及凋亡指数来评估所得囊胚的质量。添加葡萄糖增加了授精后第8天的囊胚率(80%对65%),但对孵化率没有影响(25%对28%)。在有葡萄糖的情况下观察到氧气摄取减少了22%,同时乳酸释放增加,尽管丙酮酸摄取没有变化。在有葡萄糖的情况下,第7天观察到囊胚细胞数量略有减少,而ICM/TE细胞比率和凋亡指数均未受影响。总之,从桑椹胚阶段添加5.5 mM葡萄糖对囊胚率和质量的影响有限,尽管在胚胎代谢中观察到了重要变化。这些变化是否会影响移植后胚胎的活力仍有待确定。