Effects of Glutamine, Glycine and Taurine on the Development of In Vitro Fertilized Bovine Zygotes in a Chemically Defined Medium.

Abstract

ABSTRACT. 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% O 2 : 5% CO 2 : 90% N 2 . A modified synthetic oviduct fluid medium supplemented with 20 amino acids (1 mM glutamine, essential amino acids for basal medium Eagle and nonessential 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% O 2 . -KEY WORDS: bovine embryo, culture, glutamine, glycine, taurine.

In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

“…This finding agrees with previous reports [7,8], and suggests that the developmental capacity of normally fertilized oocytes to the blastocyst stage does not differ between the two categories. However, the developmental rate of cleaved zygotes to the blastocyst stage in this study was lower than that obtained under our routine procedure (30-40%) [16]. The present culture conditions may be suboptimal, since we divided embryos into several groups according to their developmental stages at 72 hpi.…”

Effects of systemic progesterone during the early luteal phase on the availabilities of amino acids and glucose in the bovine uterine lumen

“…The relatively high abundance of Gly in the uterine lumen across species suggests that it has an important role in embryo and/or conceptus development, and this is further supported by its abundance in the fetuses of swine throughout pregnancy (Wu et al 1999). Indeed, the addition of Gly to the culture medium enhances the development of porcine and bovine embryos in vitro (Moore and Bondioli 1993;Lee and Fukui 1996;Takahashi and Kanagawa 1998;Mito et al 2012). Although the mechanisms are not well defined, Gly may serve several functions during embryo and/or conceptus development, including acting as a precursor to protein or nucleic acid synthesis, regulation of intracellular pH and osmoregulation (Edwards et al 1998;Steeves et al 2003;Tartia et al 2009;Moravek et al 2012).…”

The efficacy of the well of the well (WOW) culture system on development of bovine embryos in a small group and the effect of number of adjacent embryos on their development

“…In vitro fertilization (IVF) was conducted according to a procedure described previously (Takahashi & Kanagawa, 1998). Briefly, after the thawing of frozen semen, motile sperm were separated using a Percoll gradient (45 and 90%).…”