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Modification of ovary stock solution with magnesium and raffinose improves the developmental competence of oocytes after long preservation

Published online by Cambridge University Press:  03 January 2006

H. Iwata
Affiliation:
Department of Animal Science, Tokyo University of Agriculture, Funako 1737, Atugi 246-0034, Japan.
T. Hayashi
Affiliation:
Department of Animal Science, Tokyo University of Agriculture, Funako 1737, Atugi 246-0034, Japan.
H. Sato
Affiliation:
Department of Animal Science, Tokyo University of Agriculture, Funako 1737, Atugi 246-0034, Japan.
K. Kimura
Affiliation:
National Institute of Livestock and Grassland Science, Tochigi 329-2793, Japan.
T. Kuwayama
Affiliation:
Department of Animal Science, Tokyo University of Agriculture, Funako 1737, Atugi 246-0034, Japan.
Y. Monji
Affiliation:
Department of Animal Science, Tokyo University of Agriculture, Funako 1737, Atugi 246-0034, Japan.

Abstract

During ovary storage oocytes lose some of their developmental competence. In the present study, we maintained storage solutions of phosphate-buffered saline (PBS) at various temperatures (20 or 35 °C) or supplemented them with magnesium (Mg), raffinose and sucrose. Subsequently, we examined the kinetics of electrolytes in the follicular fluid (FF) during the ovary storage period (9h), the survival rate of granulosa cells in the follicles, and the developmental competence of oocytes after the storage. Lowering the temperature from 35 to 20 °C increased the total cell number of blastocysts that developed at 7 days after in vitro maturation and in vitro fertilization of oocytes. In stock solution with supplements of 15 mM Mg or a combination of 5 mM Mg and 10 mM raffinose or sucrose, a significantly higher number of oocytes developed into blastocysts with a large number of cells in each blastocyst, and a significantly higher number of living granulosa cells were obtained as compared with stock solutions without any supplements. During ovary storage, the concentrations of potassium and chloride in the FF were increased, and the addition of Mg to the stock solution increased the concentration of Mg in the FF. Germinal vesicle breakdown in oocytes that were collected from ovaries stored in the solution supplemented with 15 mM Mg or a combination of 5 mM Mg and 10 mM of raffinose occurred at a slower rate than that in oocytes collected from ovaries stored in PBS alone. On the other hand, the oocytes collected from ovaries stored in the solution supplemented with 15 mM Mg or a combination of 5 mM Mg and 10 mM raffinose reached the metaphase II (MII) stage more rapidly than the oocytes collected from ovaries stored in the PBS alone. In conclusion, the modification of stock solution by the addition of Mg and raffinose improved the developmental competence of oocytes obtained from ovaries preserved for a long period.

Type
Research Article
Copyright
2005 Cambridge University Press

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