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Journal of Animal Science, Vol 78, Issue 1 152-157, Copyright © 2000 by American Society of Animal Science

JOURNAL ARTICLE

Reduced oxygen tension and EDTA improve bovine zygote development in a chemically defined medium

S. E. Olson and G. E. Seidel Jr
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523, USA.

Bovine zygotes produced by in vitro oocyte maturation and fertilization were cultured for 7.5 d in a chemically defined medium without serum or proteins, except .12 IU/mL of insulin. In Exp. 1, embryos were cultured in approximately 20% oxygen (i.e., 5% CO2 in air) or 5% CO2; 5% O2; 90% N2, with the metal chelators EDTA or diethylenetetraaminopentaacetic acid (DTPA) at 0, 5, 25, or 125 microM. More (P < .01) embryos developed to blastocysts at 5% O2 (17%) than at -20% O2 (7%). Also, embryos grown at 5% O2 averaged more cells than embryos cultured at -20% O2 (38 vs 29 cells for morulae and blastocysts and 15 vs 12 cells including all embryos; P < .05). There were interactions (P < .01) among chelator, concentration of chelator, and oxygen tension. The most efficacious treatments were 5 microM EDTA at 5 or -20% O2 (24 and 20% blastocysts), 5 microM DTPA at 5% O2 (28% blastocysts), and 25 microM EDTA at 5% O2 (25% blastocysts). High concentrations of either chelator were detrimental, especially at -20% O2. In Exp. 2, a smaller range of chelator concentrations was compared (EDTA: 3, 9, 27, or 81 microM, DTPA: 3 or 15 microM) in 5% O2. More embryos developed to blastocysts and expanded blastocysts with 3 microM EDTA than the control without a chelator (20 and 16% vs 7 and 3%, respectively; P < .05). However, in Exp. 3, which concerned embryo development in .33, 1, 3, or 27 microM EDTA and .33, 1, or 3 microM DTPA, no concentration of either chelator was better (P > . 1) than the control.


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