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J Reprod Dev. 2017 Apr 21;63(2):175-183. doi: 10.1262/jrd.2016-142. Epub 2017 Feb 5.

Significant heparin effect on bovine embryo development during sexed in vitro fertilization.

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Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210046, PR China.
TransOva Genetics, Iowa 51250, USA.
Department of Animal Science, Tshwane University of Technology, Pretoria 0001, South Africa.
Renova Life, Inc., Maryland 20742, USA.
Sexing Technologies, Texas 77868, USA.
Wuxi Entry-Exit Inspection and Quarantine Bureau, Jiangsu 214101, PR China.
ARSIAL, Rome 00162, Italy.


The aim of this study was to investigate the effect of different heparin concentrations in the course of sexed in vitro fertilization (IVF), on bovine embryonic development and development to term following embryo transfer (ET). With a total of 9156 oocytes for IVF, sorted as well as unsorted sperm from four bulls had different heparin requirements for achieving the highest rate of development in vitro. However, when optimal heparin concentrations were used (40 to 80 µg/ml), the performance of X-sorted sperm (0.3 × 106/ml/IVF droplet) from all four bulls, as judged by blastocyst development (Bulls A, B, C, and D: 25.2, 19.7, 25.1, and 9.8%, respectively), was significantly increased, and the blastocyst rate was comparable to that observed with unsorted sperm at certain heparin concentrations within the four bulls. We determined that near-optimal blastocyst development was possible with sorted sperm from all four bulls, when a heparin concentration of 40 µg/ml was used. Pregnancy rates at d 70 post ET ranged from 39.1 to 40.3% (P > 0.05), and the calving rates ranged from 34.4 to 35.1% (P > 0.05), when heparin was used at a concentration of 10 μg/ml (n = 236), 20 μg/ml (n = 189), and 40 μg/ml (n = 305), respectively. Our study demonstrates that, although the sorted sperm of different bulls performed optimally over a range of heparin concentrations, a generally accepted heparin concentration of 40 µg/ml can be set for sexed IVF. This improvement is beneficial when sexed embryo production by ovum pickup and IVF is an essential component of genetic breeding programs.


Bovine; Embryo transfer; Embryos; In vitro fertilization (IVF); Sorted sperm

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