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Developmental Potential of Vitrified Holstein Cattle Embryos Fertilized In Vitro with Sex-Sorted Sperm

J. Xu^*, Z. Guo, L. Su, T. L. Nedambale^*, J. Zhang, J. Schenk^#, J. F. Moreno^||, A. Dinnyés^¶, W. Ji, X. C. Tian, X. Yang and F. Du^*,1

^* Evergen Biotechnologies, Inc., Storrs, CT 06269
Xinjiang Academy of Animal Science, the Key Laboratory of Animal Biotechnology of Ministry of Agriculture, Urumqi, Xinjiang 830000, China
Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China
Department of Animal Science/Center for Regenerative Biology, University of Connecticut, Storrs 06269
^# XY Inc., Fort Collins, CO 80524
^|| Sexing Technologies, Navasota, TX 77868
^¶ Department of Animal Biology, Agricultural Biotechnology Center, Godollo, Hungary

¹ Corresponding author: fuliangd{at}evergen.com

In vitro fertilization (IVF) is a feasible way to utilize sex-sorted sperm to produce offspring of a predetermined sex in the livestock industry. The objective of the present study was to examine the effects of various factors on bovine IVF and to systematically improve the efficiency of IVF production using sex-sorted sperm. Both bulls and sorting contributed to the variability among differential development rates of embryos fertilized by sexed sperm. Increased sorting pressures (275.8 to 344.75 kPa) did not have a significant effect on the in vitro fertility of the sorted sperm; neither did an extended period of 9 to 14 h from semen collection to sorting. As few as 600 sorted sperm were used to fertilize an oocyte, resulting in blastocyst development of 33.2%. Postwarming of vitrified sexed IVF embryos resulted in high morphological survival (96.3%) and hatching (84.4%) rates, similar to those fertilized by nonsexed sperm (93.1 and 80.6%, respectively). A 40.9% pregnancy rate was established following the transfer of 3,627 vitrified, sexed embryos into synchronized recipients. This was not different from the rates with nonsexed IVF (41.9%, n = 481), or in vivo-produced (53.1%, n = 192) embryos. Of 458 calves born, 442 (96.5%) were female and 99.6% appeared normal. These technologies (sperm sexing-IVF-vitrification-embryo transfer) provide farmers, as well as the livestock industry, with a valuable option for herd expansion and heifer replacement programs. In summary, calves were produced using embryos fertilized by sex-sorted sperm in vitro and cryopreserved by rapid cooling vitrification.

Key Words: sorted sperm • in vitro fertilization • vitrification • embryo transfer

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