Early Development of IVM/IVF Bovine Embryos Cultured with or without Somatic Cells in a Simple Serum-free Medium with Different Concentrations of CO<SUB>2</SUB> and O<SUB>2</SUB>

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概要

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• The effect of various O<SUB>2</SUB> and CO<SUB>2</SUB> mixtures on development of bovine embryos produced by <I>in vitro</I> maturation and <I>in vitro</I> fertilization (IVM/IVF) was examined in a newly formulated simple KSOM medium versus a complex Menezo B<SUB>2</SUB> serum-free medium and in co-culture. Numbers of cells comprising blastocysts were also counted using Hoechst 33342 stain. Following routine IVM/IVF procedures oocytes and zygotes were cultured for 40 to 44 h in various media. Then 2-to 8-cell embryos had cumulus cells removed and were alotted randomly for continued culture to the same experimental culture media, but with various gas conditions. In Experiment 1, the percentage of embryos developing to and beyond morula stages in 5, 10 and 20% O<SUB>2</SUB> with 5% CO<SUB>2</SUB> in humidified air was 22, 15 and 12%, respectively, with the 22 versus 12% being different (P<0.05). Blastocyst cell number in the 5 and 10% O<SUB>2</SUB> treatments was higher than in 20% O<SUB>2</SUB> (P<0.05). Also, the simple KSOM and complex Menezo B<SUB>2</SUB> media were compared and more embryos developed into blastocysts (10%) in KSOM medium than in B<SUB>2</SUB> medium (5%; P<0.05), but cell number did not differ (P>0.05). In Experiment 2, the proportions of embryos developing into blastocysts in 5 and 10% CO<SUB>2</SUB> were 15 and 6% (P<0.05), respectively, and in 5 and 10% O<SUB>2</SUB> were 15 and 7%, respectively (P<0.05). Mean blastocyst cell number (82) was highest with 5% CO<SUB>2</SUB> and 10% O<SUB>2</SUB> even though more blastocysts were obtained with 5% CO<SUB>2</SUB> and 5% O<SUB>2</SUB> (P<0.05). In Experiment 3, embryos were co-cultured on monolayers of bovine oviduct epithelial cells (BOEC) or buffalo rat liver cells with KSOM medium in 5 and 10% O<SUB>2</SUB> and 5% CO<SUB>2</SUB>. Treatment effects did not differ except 10% O<SUB>2</SUB> was superior to 5% O<SUB>2</SUB> when considering both morulae and blastocysts (41 vs 33%; P<0.05). These experiments indicate that the O<SUB>2</SUB>-CO<SUB>2</SUB> concentrations can affect the development of bovine embryos produced by IVM/IVF when cultured in a simple serum-free medium with and without co-culture and that the simple KSOM medium can serve as a defined medium to study required supplementation for culturing IVM/IVF bovine oocytes.

• 日本繁殖生物学会の論文

著者

• Foote Robert

  Department Of Animal Science Cornell University

• YANG Boo

  Department of Animal Science, Cornell University

• YANG Xiangzhong

  Department of Animal Science, Cornell University

• Yang Xiangzhong

  Department Of Animal Science University Of Connecticut

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