주요연혁 / 연구실적
연구활동 Effects of phosphate, energy substrates, and amino acids on developmen…
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Effects of phosphate, energy substrates, and amino acids on development of in vitro-matured, in vitro-fertilized bovine oocytes in a chemically defined,protein-free culture medium
JH Kim, K Niwa, JM Lim and K Okuda
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, Okayama 700, Japan.
Abstract
Bovine oocytes that had been matured and fertilized in vitro were cultured in a simple, chemically defined, protein-free medium (mTLP- PVA). When the medium was supplemented with 19 amino acids,
development to the 8-cell (14-20% vs. 38-46%), morula (0-6% vs. 27-32%), and blastocyst (0-1% vs. 9-13%) stages 96, 144, and 192 h after insemination, respectively, was significantly greater in the
absence than in the presence of glucose (5.56 mM) regardless of the presence of phosphate (1.05 mM). However, blastocyst development was difficult in medium with any combination of glucose and phosphate
without amino acids. In mTLP-PVA with amino acids and different concentrations of phosphate, the highest proportions of embryos reaching the > or = 8- cell (56%), morula (44%), and blastocyst (24%) stages
were obtained at a 0.35 mM concentration. When lactate and pyruvate were omitted from mTLP-PVA (mT-PVA) supplemented with amino acids and 0.35 mM phosphate, the first cleavage was completely
inhibited. Although lactate or pyruvate alone could support blastocyst development to a limited extent (10-15%), a significantly higher proportion (22%) of blastocysts was obtained in medium with both lactate (10
mM) and pyruvate (0.5 mM).