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 Jong-Heung KIm, Koji NIWA, Jeong-Mook LIM, Kiyoshi OKUDA 

​Biology of Reproduction 48:1320-1325, 1993 

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).