Abstract
In vitro embryo production using oocytes of prepubertal females is named JIVET (juvenile in vitro embryo technologies). The aim of the JIVET is to increase genetic gain by shortening the generation interval in breeding programs. In this chapter we describe the methodology currently used in our laboratory for producing in vitro embryos from prepubertal sheep (90–120 days old) and goats (30–45 days old).
Briefly, we obtain cumulus–oocyte complexes (COCs) from slaughtered female ovaries. These COCs are in vitro matured in TCM199 with 10% (v/v) fetal bovine serum (FBS) at 38.5 °C in a humidified air atmosphere with 5% CO2 for 24 h. The next day we collect ejaculates from males of proven fertility and select the most motile spermatozoa by density gradient. Following coincubation with mature COCs, presumptive zygotes and embryos are cultured in SOF for 8 days at 38.5 °C, 90% N2, 5% CO2, 5% O2, and 100% humidity to obtain blastocysts.
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References
Morton KM (2008) Developmental capabilities of embryos produced in vitro from prepubertal lamb oocytes. Reprod Domest Anim 43:137–143
Koeman J, Keefer CL, Baldassarre H, Downey BR (2003) Developmental competence of prepubertal and adult goat oocytes cultured in semi-defined media following laparoscopic recovery. Theriogenology 60:879–889
Paramio MT, Izquierdo D (2016) Recent advances in in vitro embryo production in small ruminants. Theriogenology 86:152–159
Armstrong DT (2001) Effects of maternal age on oocyte developmental competence. Theriogenology 55:1303–1322
Paramio MT, Izquierdo D (2014) Current status of in vitro embryo production in sheep and goats. Reprod Domest Anim 249:37–48
Velilla E, Izquierdo D, Rodriguez-Gonzalez E et al (2004) Distribution of prepubertal and adult goat oocyte cortical granules during meiotic maturation and fertilisation: ultrastructural and cytochemical study. Mol Reprod Dev 68:507–514
Velilla E, Rodriguez-Gonzalez E, Vidal F et al (2006) Mitochondrial organization in prepubertal goat oocytes during in vitro maturation and fertilization. Mol Reprod Dev 73:617–626
Anguita B, Jiménez-Macedo AR, Izquierdo D et al (2007) Effect of oocyte diameter on meiotic competence, embryo development, p34 (cdc2) expression and MPF activity in prepubertal goat oocytes. Theriogenology 67:526–536
Anguita B, Paramio M, Jiménez-Macedo A et al (2008) Total RNA and protein content, Cyclin B1 expression and developmental competence of prepubertal goat oocytes. Anim Reprod Sci 103:290–303
Velilla E, Rodríguez-González E, Vidal F, Paramio MT (2005) Microtubule and microfilament organization in immature, in vitro matured and in vitro fertilized prepubertal goat oocytes. Zygote 13:55–165
Villamediana P, Vidal F, Paramio MT (2001) Cytogenetic analysis of caprine 2- to 4-cell embryos produced in vitro. Zygote 9:193–199
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Izquierdo, D., Catalá, MG., Paramio, MT. (2019). Small Ruminants: Prepubertal Oocyte Donors. In: Herrick, J. (eds) Comparative Embryo Culture. Methods in Molecular Biology, vol 2006. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9566-0_11
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DOI: https://doi.org/10.1007/978-1-4939-9566-0_11
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Publisher Name: Humana, New York, NY
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