Takahashi Y, Kanagawa H
Department of Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
J Vet Med Sci. 1998 Apr;60(4):433-7. doi: 10.1292/jvms.60.433.
The effects of glutamine, glycine and taurine on the development of bovine zygotes derived from IVM/IVF were determined using a protein-free chemically defined medium. After the cumulus cells were removed at 18 hr post-insemination, the presumptive zygotes were cultured for 4 or 6 days (about 104 or 154 hr) under a gas atmosphere of 5% O2: 5% CO2: 90% N2. A modified synthetic oviduct fluid medium supplemented with 20 amino acids (1 mM glutamine, essential amino acids for basal medium Eagle and non-essential amino acids for minimum essential medium), insulin, and PVA was used as a basic medium (mSOFai). Omitting 1 mM glutamine from mSOFai did not affect the embryonic development after 4 and 6 days of culture. After 4 days of culture, no significant effects of glycine and taurine on the development of zygotes to the morula stage were observed. However, supplementation with glycine or taurine significantly (P < 0.05) affected, with no interaction, the embryonic development to blastocysts after 6 days of culture. Addition of 5 mM glycine and 2 or 10 mM taurine significantly (P < 0.05) increased the percentage of blastocysts. The mean cell number in the blastocysts was affected by the glycine level, and was increased by the addition of 10 mM glycine (P < 0.001). These results demonstrate that glycine and taurine in a chemically defined medium containing a group of essential and non-essential amino acids improve the development of bovine zygotes to the blastocyst stage under 5% O2.
使用无蛋白化学限定培养基,测定了谷氨酰胺、甘氨酸和牛磺酸对体外成熟/体外受精获得的牛受精卵发育的影响。在授精后18小时去除卵丘细胞,将推定的受精卵在5% O₂: 5% CO₂: 90% N₂的气体氛围下培养4天或6天(约104或154小时)。一种添加了20种氨基酸(1 mM谷氨酰胺、伊格尔基础培养基的必需氨基酸和最低必需培养基的非必需氨基酸)、胰岛素和聚乙烯醇的改良合成输卵管液培养基用作基础培养基(mSOFai)。从mSOFai中省略1 mM谷氨酰胺对培养4天和6天后的胚胎发育没有影响。培养4天后,未观察到甘氨酸和牛磺酸对受精卵发育至桑椹胚阶段有显著影响。然而,添加甘氨酸或牛磺酸对培养6天后胚胎发育至囊胚阶段有显著(P < 0.05)影响,且无相互作用。添加5 mM甘氨酸和2或10 mM牛磺酸显著(P < 0.05)提高了囊胚百分比。囊胚中的平均细胞数受甘氨酸水平影响,添加10 mM甘氨酸可使其增加(P < 0.001)。这些结果表明,在含有一组必需和非必需氨基酸的化学限定培养基中,甘氨酸和牛磺酸可改善牛受精卵在5% O₂条件下发育至囊胚阶段的情况。