TY - CHAP
T1 - Small Ruminants: Prepubertal Oocyte Donors
AU - Izquierdo, Dolors
AU - Catalá, María Gracia
AU - Paramio, María Teresa
PY - 2019/1/1
Y1 - 2019/1/1
N2 - © Springer Science+Business Media, LLC, part of Springer Nature 2019. 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.
AB - © Springer Science+Business Media, LLC, part of Springer Nature 2019. 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.
KW - Embryo
KW - Goat
KW - In vitro culture
KW - In vitro fertilization
KW - In vitro maturation
KW - Juvenile in vitro embryo technology
KW - Oocyte
KW - Prepubertal
KW - Sheep
UR - http://www.mendeley.com/research/small-ruminants-prepubertal-oocyte-donors
U2 - 10.1007/978-1-4939-9566-0_11
DO - 10.1007/978-1-4939-9566-0_11
M3 - Chapter
C2 - 31230279
SN - 1064-3745
VL - 2006
T3 - Methods in Molecular Biology
SP - 155
EP - 163
BT - Methods in Molecular Biology
ER -