Nuclear maturation and development of IVM/IVF canine embryos in synthetic oviductal fluid or in co-culture with buffalo rat liver cells

Saikhun, J.; Sriussadaporn, S.; Thongtip, N.; Pinyopummin, A.; Kitiyanant, Y.

doi:10.1016/j.theriogenology.2008.01.024

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Volume 69, Issue 9 , Pages 1104-1110, June 2008

Nuclear maturation and development of IVM/IVF canine embryos in synthetic oviductal fluid or in co-culture with buffalo rat liver cells

J. Saikhun
- Institute of Science and Technology for Research and Development, Mahidol University, Puthamonthon sai 4, Salaya, Nakhonpathom 73170, Thailand
- Corresponding author. Tel.: +662 4419003–7x1389/90; fax: +662 4411013.
S. Sriussadaporn
- Center for Agricultural Biotechnology, Kasetsart University, Kamphaengsaen Campus, Nakhonpathom 73140, Thailand
N. Thongtip
- Faculty of Veterinary Medicine, Kasetsart University, Kamphaengsaen Campus, Nakhonpathom 73140, Thailand
A. Pinyopummin
- Center for Agricultural Biotechnology, Kasetsart University, Kamphaengsaen Campus, Nakhonpathom 73140, Thailand
- Faculty of Veterinary Medicine, Kasetsart University, Kamphaengsaen Campus, Nakhonpathom 73140, Thailand
Y. Kitiyanant
- Institute of Science and Technology for Research and Development, Mahidol University, Puthamonthon sai 4, Salaya, Nakhonpathom 73170, Thailand
- Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand

Received 4 November 2007; received in revised form 3 January 2008; accepted 29 January 2008. published online 26 March 2008.

Abstract

In vitro embryo production in the domestic bitch can provide valuable insights for conservation of endangered canids. In the present study, canine oocytes underwent in vitro maturation (IVM) in simple or complex media, with production of in vitro matured and fertilized (IVM/IVF) canine embryos. Cumulus–oocyte complexes (COCs) were harvested from ovaries by slicing and subjected to IVM in four media (SOF, TCM 199, Ham-F10, and DMEM/F12). After culture for 48h, oocytes were stained and examined for nuclear maturation. There were no significant differences in the mean (±S.D.) percentage of nuclear maturation (metaphase II) of oocytes cultured in SOF (18.6±7.6%), TCM 199 (18.3±4.5%), Ham-F10 (13.9±8.2%), or DMEM/F12 (11.9±4.2%). For assessment of embryo development, oocytes were matured for 48h in synthetic oviductal fluid (SOF), fertilized with frozen-thawed sperm, and presumptive zygotes were cultured for 7 d, either in SOF or as co-cultures with BRL cells in TCM 199. Percentages of IVM/IVF oocytes that developed to the 2-cell, 3–4-cell, and 5–7-cell stages were higher (P<0.05) following culture in SOF versus BRL cell co-cultures (33.6±1.2% vs 13.7±1.2%, 24.7±0.5% vs 8.7±1.1%, and 15.1±2.2% vs 4.3±1.3%, respectively). However, none of the embryos developed beyond the 8–16-cell stage. In conclusion, simple or complex media successfully induced resumption of meiosis and nuclear maturation of canine oocytes. Furthermore, SOF supported in vitro development of IVM/IVF canine embryos to the 8–16-cell stage.