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

• Published in: Theriogenology 2008-06, Vol.69 (9), p.1104-1110
• Main Authors: Saikhun, J., Sriussadaporn, S., Thongtip, N., Pinyopummin, A., Kitiyanant, Y.
• Format: Article
• Language: English
• Subjects: Animals; Body Fluids; Canine; cell nucleus; coculture; Coculture Techniques; culture media; cumulus oophorus; Dog; dogs; Dogs - embryology; embryo (animal); embryo culture; embryogenesis; Embryonic Development; Fallopian Tubes - physiology; Female; females; Fertilization in Vitro - veterinary; hepatocytes; In vitro fertilization; In vitro maturation; laboratory techniques; Liver - cytology; meiosis; Oocyte; Oocytes; ovaries; oviductal fluids; Rats; Rats, Inbred BUF; Semen; synthetic oviductal fluids; zygote
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/j.theriogenology.2008.01.024

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 48 h, 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 48 h 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.