Prematurational Culture with 3-Isobutyl-1-methylxanthine Synchronizes Meiotic
Progression of the Germinal Vesicle Stage and Improves Nuclear Maturation and Embryonic
Development in In Vitro-grown Bovine Oocytes |
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Authors: | Weiping HUANG Masashi NAGANO Sung-Sik KANG Yojiro YANAGAWA Yoshiyuki TAKAHASHI |
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Institution: | 1) Laboratory of Theriogenology, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan |
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Abstract: | The objective of this study was to clarify the effects of prematurational culture
(pre-IVM) supplemented with 3-isobutyl-1-methylxanthine (IBMX) on nuclear and cytoplasmic
maturation of in vitro-grown bovine oocytes. In experiment 1, oocytes (95
μm in diameter) derived from early antral follicles (0.5–1 mm in diameter) were cultured
for 12 days for in vitro growth (IVG). IVG oocytes with a normal
appearance were subjected to examinations of diameter and chromatin structure in the
germinal vesicle (GV) before IVM. In addition, percentages of metaphase II (M II) were
examined after IVM. Regardless of pre-IVM, the mean diameters of IVG oocytes were about
115 μm. The proportions of GV3 (50.0%) and M II stages (80.1%) of IVG oocytes with pre-IVM
were higher than those without pre-IVM (28.0 and 49.4%, respectively). In experiment 2,
the fertilizability and developmental competence of IVG oocytes were examined. Regardless
of pre-IVM, the normal fertilization rates of IVG oocytes were similar (around 70%) but
were lower than that of in vivo-grown oocytes (88.0%). Cleavage and
blastocyst rates of IVG oocytes with pre-IVM (63.0 and 26.1%, respectively) were higher
than those without pre-IVM (45.8 and 12.7%, respectively). The blastocyst rate based on
cleaved IVG oocytes with pre-IVM (41.7%) was similar to that of in
vivo-grown oocytes (48.7%), although the cleavage rate of IVG oocytes with
pre-IVM was lower than that of in vivo-grown oocytes. In conclusion,
pre-IVM with IBMX improved the maturational and developmental competences of IVG oocytes,
probably due to promotion of their chromatin transition and synchronization of meiotic
progression. |
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Keywords: | Early antral follicle Fertilization Germinal vesicle IBMX Pre-IVM culture |
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