Optimization of a Bovine Intracytoplasmic Sperm Injection System Using Sex-Sorted Sperm: Screening with Slaughterhouse-Derived Oocytes and Validation with Ovum Pick-Up-Derived Oocytes.
- 1Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Xianyang 712100, China.
- 2Key Laboratory of Animal Biotechnology of the Ministry of Agriculture and Rural Affairs, Northwest A&F University, Xianyang 712100, China.
Abstract
Sex-sorted sperm permit the preferential production of offspring of a specified sex, but flow sorting and cryopreservation can compromise sperm function and reduce the efficiency of conventional in vitro fertilization (IVF). We sequentially evaluated oocyte maturation duration, sperm processing, post-injection incubation medium, and nine activation regimens using slaughterhouse-derived oocytes, and then evaluated the best-performing combination under the tested sequential conditions using ovum pick-up (OPU)-derived oocytes. In conventional IVF, the presumptive normal fertilization rate, based on the presence of two pronuclei (2PN), was 2.45% with X-sorted sperm and 46.13% with conventional sperm. The final selected combination, consisting of 22 h maturation, Brackett-Oliphant (BO)-based swim-up, oocyte maturation medium (OM) during post-injection incubation, and treatment with 7% ethanol for 5 min [ETH (5)], resulted in a presumptive normal fertilization (2PN) rate of 52.92%, a blastocyst rate of 10.46%, and an ICM/TE ratio of 0.82. In the validation using OPU-derived oocytes, the blastocyst formation rate after ssICSI was 13.46% when calculated relative to injected oocytes and 16.84% when calculated relative to surviving oocytes. These rates differed significantly from the blastocyst formation rate after ssIVF (7.88%), whereas cleavage rates did not differ significantly between groups. ETH (5) provided the most favorable in vitro balance of presumptive 2PN formation, blastocyst development, and ICM/TE cell allocation among the tested regimens. Because the factors were optimized sequentially, their interactions were not estimated, which represents a limitation of this study. Furthermore, implantation, pregnancy, and calving rates require validation in independent in vivo embryo-transfer experiments.
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BibTeX
@article{song2026optimization,
title = {{Optimization of a Bovine Intracytoplasmic Sperm Injection System Using Sex-Sorted Sperm: Screening with Slaughterhouse-Derived Oocytes and Validation with Ovum Pick-Up-Derived Oocytes.}},
author = {Xuexiao Song and Tao Zhang and Hongxing Yang and Heqiang Li and Baobao Zhao and Yanfang Wu and Zheng Wang and Dexiang Fan and Yaochang Wei and Dong Zhou and Yongsheng Wang},
journal = {Animals : an open access journal from MDPI},
year = {2026},
month = sep,
doi = {10.3390/ani16192983},
url = {https://europepmc.org/article/MED/42882952},
note = {CC BY},
}
RIS
TY - JOUR
TI - Optimization of a Bovine Intracytoplasmic Sperm Injection System Using Sex-Sorted Sperm: Screening with Slaughterhouse-Derived Oocytes and Validation with Ovum Pick-Up-Derived Oocytes.
AU - Xuexiao Song
AU - Tao Zhang
AU - Hongxing Yang
AU - Heqiang Li
AU - Baobao Zhao
AU - Yanfang Wu
AU - Zheng Wang
AU - Dexiang Fan
AU - Yaochang Wei
AU - Dong Zhou
AU - Yongsheng Wang
PY - 2026
DA - 2026/09/22
JO - Animals : an open access journal from MDPI
DO - 10.3390/ani16192983
UR - https://europepmc.org/article/MED/42882952
AB - Sex-sorted sperm permit the preferential production of offspring of a specified sex, but flow sorting and cryopreservation can compromise sperm function and reduce the efficiency of conventional in vitro fertilization (IVF). We sequentially evaluated oocyte maturation duration, sperm processing, post-injection incubation medium, and nine activation regimens using slaughterhouse-derived oocytes, and then evaluated the best-performing combination under the tested sequential conditions using ovum pick-up (OPU)-derived oocytes. In conventional IVF, the presumptive normal fertilization rate, based on the presence of two pronuclei (2PN), was 2.45% with X-sorted sperm and 46.13% with conventional sperm. The final selected combination, consisting of 22 h maturation, Brackett-Oliphant (BO)-based swim-up, oocyte maturation medium (OM) during post-injection incubation, and treatment with 7% ethanol for 5 min [ETH (5)], resulted in a presumptive normal fertilization (2PN) rate of 52.92%, a blastocyst rate of 10.46%, and an ICM/TE ratio of 0.82. In the validation using OPU-derived oocytes, the blastocyst formation rate after ssICSI was 13.46% when calculated relative to injected oocytes and 16.84% when calculated relative to surviving oocytes. These rates differed significantly from the blastocyst formation rate after ssIVF (7.88%), whereas cleavage rates did not differ significantly between groups. ETH (5) provided the most favorable in vitro balance of presumptive 2PN formation, blastocyst development, and ICM/TE cell allocation among the tested regimens. Because the factors were optimized sequentially, their interactions were not estimated, which represents a limitation of this study. Furthermore, implantation, pregnancy, and calving rates require validation in independent in vivo embryo-transfer experiments.
SN - 2076-2615
ER -