Rosenkrans C F, Zeng G Q, MCNamara G T, Schoff P K, First N L
Department of Meat and Animal Science, University of Wisconsin, Madison 53706.
Biol Reprod. 1993 Sep;49(3):459-62. doi: 10.1095/biolreprod49.3.459.
Simple media were developed to study the metabolic requirements of bovine embryos up to Day 7 (Day 0 = day of oocyte aspiration) in vitro. Embryos were derived from oocytes matured and fertilized in vitro. At 45 +/- 2 h post insemination, embryos (> or = 2 cells) were randomly allotted to treatments. Examined in experiments 1 and 3 was the effect of pyruvate concentration in the presence of lactate. In the presence of lactate, pyruvate (0.2-5.0 mM) had no effect (p > 0.05) on the percentage of morulae or blastocysts. However, increasing the concentration of hemicalcium L-lactate from 5 mM to 10 mM decreased (p < 0.001) the percentage of embryos reaching the morula or blastocyst stage (experiment 3). Neither magnesium sulfate (0.5 mM) nor EDTA (10 mM) improved embryo development when added to the medium CR1 (experiment 2). Increasing the calcium level to 5 mM or the lactate level to 10 mM had no effect (p > 0.05) on embryo development (experiment 4). However, the interaction of adding calcium and lactate resulted in a decreased (p < 0.05) percentage of morulae. Determined in experiment 6 were the independent effects of pyruvate, lactate, and glucose on embryo development in vitro. As pyruvate or lactate level was increased from 1 to 10 mM, the percentage of blastocysts was decreased (p < 0.05). These experiments indicate that adding pyruvate to a medium containing lactate is not necessary for development of bovine embryos in vitro.
开发了简单培养基以研究体外培养至第7天(第0天 = 卵母细胞抽吸日)的牛胚胎的代谢需求。胚胎来源于体外成熟和受精的卵母细胞。在授精后45±2小时,将胚胎(≥2细胞)随机分配至各处理组。在实验1和3中研究了乳酸存在时丙酮酸浓度的影响。在乳酸存在的情况下,丙酮酸(0.2 - 5.0 mM)对桑葚胚或囊胚的百分比没有影响(p>0.05)。然而,将L - 乳酸半钙的浓度从5 mM增加到10 mM会降低(p<0.001)达到桑葚胚或囊胚阶段的胚胎百分比(实验3)。当添加到CR1培养基中时,硫酸镁(0.5 mM)和EDTA(10 mM)均未改善胚胎发育(实验2)。将钙水平提高到5 mM或乳酸水平提高到10 mM对胚胎发育没有影响(p>0.05)(实验4)。然而,添加钙和乳酸的相互作用导致桑葚胚百分比降低(p<0.05)。在实验6中确定了丙酮酸、乳酸和葡萄糖对体外胚胎发育的独立影响。随着丙酮酸或乳酸水平从1 mM增加到10 mM,囊胚的百分比降低(p<0.05)。这些实验表明,向含有乳酸的培养基中添加丙酮酸对于牛胚胎的体外发育不是必需的。