Vet Med - Czech, 2010, 55(1):10-18 | DOI: 10.17221/160/2009-VETMED

Conception rate after sex determination and cryopreservation of D7 bovine embryos:

M. Lopatarova1, S. Cech1, P. Krontorad2, L. Holy1, H. Lalova1, R. Dolezel1
1 University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic
2 BOVET Company, Sloupnice, Czech Republic

The aim of this study was to evaluate conception rate after sex determination and subsequent freezing of Day 7 (D7) bovine embryos and to compare it with transfer of fresh female embryos. High quality embryos obtained from superovulated donors were biopsied and the embryonic cells (5-10) were analysed by PCR using specific primers for the Y-chromosome determinant. Fresh embryos were blade biopsied (n = 172) and subsequently transferred (ET) ipsilaterally to synchronized recipients. Selected embryos (n= 112) were biopsied by cell aspiration and cryopreserved for later transfer. Sex determination was successfully completed in 91.3% of fresh group embryos (44% female) and in 87.5% of freeze-thawed group embryos (45.9% female). The achieved pregnancy rate was 56.5% after fresh transfer of sexed (female) embryos whereas intact embryos (control 1) implanted in 61.9% recipients. 95.6% (43/45) of embryos survived biopsy and cryopreservation. After transfer of freeze-thawed sexed (female) embryos, pregnancy was established in 48.8% of recipients. A similar conception rate was found in intact embryos (50.7%, control 2). The results clearly demonstrate that the microsurgical technique used and subsequent PCR sex analysis allow the rapid commercial exchange of genetic resources on the basis of fresh or frozen sex-desired embryos in embryo transfer programs.

Keywords: cattle; superovulation; sexing; polymerase chain reaction; cryopreservation; embryo transfer

Published: January 31, 2010  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Lopatarova M, Cech S, Krontorad P, Holy L, Lalova H, Dolezel R. Conception rate after sex determination and cryopreservation of D7 bovine embryos:. Vet Med - Czech. 2010;55(1):10-18. doi: 10.17221/160/2009-VETMED.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Alonso RV, Hellu JAA, Perri SHV, Visintin JA, Garcia JF (2009): Factors affecting commercial sexing program and multiple genetic analysis perspectives of in vitroproduced bovine embryos. Reproduction, Fertility and Development, 21, 227. Go to original source...
    2. Andersson M, Taponen J, Kommeri M, Dahlbom M (2006): Pregnancy rates in lactating Hostein-Friesian cows after artificial insemination with sexed sperm. Reproduction in Domestic Animals, 41, 95-97. Go to original source... Go to PubMed...
    3. Avery B, Bak A, Schmidt M (1989): Different cleavage rates and sex determination in bovine embryos. Theriogenology, 32, 139-147. Go to original source... Go to PubMed...
    4. Blondin P, Beaulieu M, Fournier V, Morin N, Crawford L, Madan P, King WA (2009): Analysis of bovine sexed sperm for IVF from sorting to the embryo. Theriogenology, 71, 30-38. Go to original source... Go to PubMed...
    5. Bondioli KR, Ellis SB, Pryor JH, Williams MW, Harpold MM (1989): The use of male-specific chromosomal DNA fragments to determine the sex of bovine preimplantation embryos. Theriogenology, 31, 95-104. Go to original source...
    6. Booman P, Kruijt L, Veerhuis R, Hengst AM, Tiemann M, Ruch FE (1989): Sexing bovine embryos with monoclonal antibodies against the H-Y antigen. Livestock Production Science (Niederlande), 23, 1-16. Go to original source...
    7. DeJarnette JM, Nebel RL, Marshall CE (2009): Evaluating the success of sex-sorted semen in US dairy herds from on farm records. Theriogenology, 71, 49- 58. Go to original source... Go to PubMed...
    8. Edwards RG, Gardner RL (1967): Sexing of live rabbit blastocyst. Nature, 214, 576- 577. Go to original source... Go to PubMed...
    9. Ekici H, Turan N, Sontas BH, Helps CR, Senunver A, Ylmaz H (2006): Sex determination of bovine embryos using polymerase chain reaction (PCR). Revue de Medicine Veterinaire, 157, 441-444.
    10. Garner DL, Seidel GE Jr. (2008): History of commercializing sexed semen for cattle. Theriogenology, 69, 886-895. Go to original source... Go to PubMed...
    11. Gutierrez-Adan A, Lonergan P, Rizos D, Ward FA, Boland MP, Pintado B, de la Fuente J (2001): Effect of the in vitro culture system on the kinetics of blastocyst development and sex ratio of bovine embryos. Theriogenology, 55, 1117-1126. Go to original source... Go to PubMed...
    12. Han Y, Yoo O, Lee K (1993): Sex determination in single mouse blastomeres by polymerase chain reaction. Journal of Assisted Reproduction and Genetics, 10, 151. Go to original source... Go to PubMed...
    13. Handyside AH, Kontogiann EH, Hardy K, Winston RML (1990): Pregnancies from biopsied human preimplantation embryos sexed by Y-specific DNA amplification. Nature, 344, 768-770. Go to original source... Go to PubMed...
    14. Hare WCD, Mitchell D, Betteridge KJ, Eaglesome MD, Randalu GCB (1976): Sexing two week-old bovine embryos by chromosomal analysis prior to surgical transfer: preliminary methods and results. Theriogenology, 5, 243-253. Go to original source...
    15. Hasler JF, Cardey E, Stokes JE., Bredbacka P (2002): Nonelectrophoretic PCR-sexing of bovine embryos in a commercial environment. Theriogenology, 58, 1457-1469. Go to original source... Go to PubMed...
    16. Hayakawa H, Hirai T, Takimoto A, Ideta A, Aoyagi Y (2009): Superovulation and embryo transfer in Holstein cattle using sexed sperm. Theriogenology, 71, 68-73. Go to original source... Go to PubMed...
    17. Herr CM, Holt NA, Matthaei KI, Reed KC (1990): Sex of progeny from bovine embryos sexed with a rapid Ychromosome-detection assay. Theriogenology, 33, 247. Go to original source...
    18. Herr CM, Steel T, Lascelles A, Reed KC (1995): Instruction Manual: Splitting, Biopsy & Rapid Sexing of Cattle Embryos. AB Technology, Inc., Pullmann, WA. 58 pp.
    19. Hirayama H, Kageyama S, Moriyasu S, Sawai K, Onoe S, Takahashi Y, Katagiri S, Toen K, Watanabe K, Notomi T, Yamashina H, Matzuzaki S, Minamihashi A (2004): Rapid sexing of bovine preimplantation embryo using loop-mediated isothermal amplification. Theriogenology, 62, 887-896. Go to original source... Go to PubMed...
    20. Hirayama H, Fujikawa A, Kageyama S, Moriyasu S, Sawai K, Onoe S, Minamihashi A (2008): Multiple genotyping in bovine pre-implantation embryo with whole genome amplification. Animal Science Journal, 79, 554-560. Go to original source...
    21. Holy L, Lopatarova M, Krontorad P, Holy J (1990): Systematic use of embryo transfer in cattle under condition of high animal concentration (in German). Monatsheft Veterinaermedizin, 45, 409-413.
    22. Ipate I, Bogdan AT, Toba LG, Toba GF., Ivana S (2007): Research regarding the embryo biopsy for sexing and genetic testing the bovines embryos. Revista Romana de Medicina Veterinara, 17, 213-218.
    23. Katska-Ksiazkiewicz L, Rynska B, Bochenek M, Opiela J, Jurkiewicz J (2006): In vitro production of bovine embryos using flow-cytometrically sexed sperm. Archiv fur Tierzucht, 49, 133-140. Go to original source...
    24. Kimura K, Iwata H, Thompson JG (2008): The effect of glucosamine concentration on the development and sex ratio of bovine embryo. Animal Reproduction Science, 103, 228-238. Go to original source... Go to PubMed...
    25. King WA, Picard L, Bosquet D, Goff AK (1992): Sexdependent loss of bisected bovine morulae after culture and freezing. Journal of Reproduction and Fertility, 96, 453-459. Go to original source... Go to PubMed...
    26. Kobayashi J, Sekimoto A, Uchida H, Wada T, Sasaki K, Sasada H, Umezu M, Sato E (1998): Rapid detection of male-specific DNA sequence in bovine embryos using fluorescence in situ hybridization. Molecular Reproduction and Development, 51, 390-394. Go to original source...
    27. Lacaze S, Humblot P, Ponsart C (2008): Sexing and direct transfer of bovine biopsied frozen-thawed embryos under on-farm conditions in a commercial program. Recherches sur les Ruminants, 15eme Recontre, Paris, les 3 et 4 decembre 2008, 387-390.
    28. Leonhard M, Korszenbaum M, Cotinot C, Chesne P, Heyman Y, Stinnakre MG, Bishop C, Delouis C, Vaiman M, Fellous M (1987): Sexing bovine embryos using Y chromosome specific DNA probe. Theriogenology, 27, 248. Go to original source...
    29. Li S, Yu W, Fu J, Bai Y, Jin F, Shangguan B (2007): Factors influencing pregnancy rates following transfer of bovine in vivo embryos biopsied for sex determination. Reproduction, Fertility and Development, 19, 297. Go to original source...
    30. Lopatarova M, Cech S, Krontorad P, Holy L, Hlavicova J, Dolezel R (2007): Lower quality bovine embryos may be successfully used for sex determination. Veterinarni Medicina, 52, 540-546. Go to original source...
    31. Lopatarova M, Cech S, Krontorad P, Holy L, Hlavicova J, Dolezel R (2008): Sex determination in bisected bovine embryos and conception rate after the transfer of female demi-embryos. Veterinarni Medicina, 53, 595-603. Go to original source...
    32. Lopes RFF, Forell F, Oliveira ATD, Rodrigues JL (2001): Splitting and biopsy for bovine embryo sexing under field conditions. Theriogenology, 56, 1383-1392. Go to original source... Go to PubMed...
    33. Monk M, Handyside AH (1988): Sexing of preimplantation mouse embryos by measurement of X-linked gene dosage in a single blastomere. Journal of Reproduction and Fertility, 82, 365-368. Go to original source... Go to PubMed...
    34. Nedambale TL, Dinnyes A, Yank XZ, Tian XC (2004): Bovine blastocyts development in vitro: timing, sex, and viability folloving vitrification. Biology of Reproduction, 71, 1671-1676. Go to original source... Go to PubMed...
    35. Peippo J, Huhtinen M, Kotilainen T (1995): Sex diagnosis of equine preimplantation embryos using the polymerase chain reaction. Theriogenology, 44, 619-627. Go to original source... Go to PubMed...
    36. Peippo J, Viitala S, Virta J, Raty M, Tammiranta N, Lamminen T, Aro J, Myllymaki H, Vilkki J (2007): Birth of correctly genotyped calves after multiplex marker detection from bovine embryo microblade biopsie. Molecular Reproduction and Development, 74, 1373-1378. Go to original source... Go to PubMed...
    37. Peippo J, Vartia K, Kananen-Anttila K, Raty M, Korhonen K, Hurme T, Myllymaki H, Sairanen A, MakiTanila A (2009): Embryo production from superovulated Holstein-Friesian dairy heifers and cows after insemination with frozen-thawed sex-sorted X spermatozoa or unsorted semen. Animal Reproduction Science, 111, 80-92. Go to original source... Go to PubMed...
    38. Pereira RM, Carvalhais I, Pimenta J, Baptista MC, Vasques MI, Horta AEM, Santos IC, Marques MR, Reis A, Silva Pereira M, Marques CC (2008): Biopsied and vitrified bovine embryos viability is improved by trans10, cis12 conjugated linoleic acid supplementation during in vitro embryo culture. Animal Reproduction Science, 106, 322-332. Go to original source... Go to PubMed...
    39. Peura T, Hyttinen JM, Turunen M, Janne J (1991): A reliable sex determination assay for bovine preimplantation embryos using the polymerase chain reaction. Theriogenology, 35, 547-555. Go to original source... Go to PubMed...
    40. Pomp P, Good BA, Geisert RD, Corbin CJ, Conley AJ (1995): Sex identification in mammals with polymerase chain reaction and its use to examine sex effect on diameter of day-10 or -11 pig embryos. Journal of Animal Science, 73, 1408-1415. Go to original source... Go to PubMed...
    41. Rall WF, Leibo SP (1987): Production of sexed bovine pregnancies by cytogenetic analysis of cultured demiembryos. Theriogenology, 27, 269. Go to original source...
    42. Robertson I, Nelson R (1998): Certification and identification of the embryo. 103-134. In: Strinkfellow D.A., Seidel S.M. (eds.): Manual of the International Embryo Transfer Society. IETS, Illinois. 171 pp.
    43. Sartori R, Souza AH, Guenther JN, Caraviello DZ, Geiger LN, Schenk JL (2004): Fertilization rate and embryo quality in superovulated Holstein heifers artificially inseminated with X-sorted or unsorted sperm. Animal Reproduction, 1, 86-90.
    44. Schenk JL, Suh TK, Seidel GE Jr. (2006): Embryo production from superovulated cattle following insemination of sexed sperm. Theriogenology, 65, 299-307. Go to original source... Go to PubMed...
    45. Schenk JL, Cran DG, Everett RW, Seidel GE Jr. (2009): Pregnancy rates in heifers and cows with cryopreserved sexed sperm: Effects of sperm numbers per inseminate, sorting pressure and sperm storage before sorting. Theriogenology, 71, 717-728. Go to original source... Go to PubMed...
    46. Schroder A, Roschlau D, Giehm D, Schwerin M, Thomsen PD (1990): Sex determination of bovine embryos using Y-specific primers in the polymerase chain reaction. Archiv fur Tierzucht, 33, 293-299.
    47. Seidel GE Jr. (2007): Overview of sexing sperm. Theriogenology, 68, 443-446. Go to original source... Go to PubMed...
    48. Shea BF (1999): Determining the sex of bovine embryos using polymerase chain reaction results: a six-year retrospective study. Theriogenology, 51, 841-854. Go to original source... Go to PubMed...
    49. Singh EL, Hare WCD (1980): The feasibility of sexing bovine morula stage embryos prior to embryo transfer. Theriogenology, 14, 421-427. Go to original source...
    50. Thibier M, Nibart M (1995): The sexing of bovine embryos in the field. Theriogenology, 43, 71-80. Go to original source...
    51. Tominaga K (2004): Cryopreservation and sexing of in vivo- and in vitro-produced bovine embryos for their practical use. Journal of Reproduction and Development, 50, 29-38. Go to original source... Go to PubMed...
    52. Tominaga K, Hamada Y (2004): Efficient production of sex-identified and cryosurvived bovine in-vitro produced blastocysts. Theriogenology, 61, 1181-1191. Go to original source... Go to PubMed...
    53. Tominaga K, Iwaki F, Hochi S (2007): Conventional freezing of in vitro-produced and biopsied bovine blastocysts in the presence of a low concentration of glycerol and sucrose. Journal of Reproduction and Development, 53, 443-447. Go to original source... Go to PubMed...
    54. Voelkel SA, Hu YX (1992): Direct transfer of frozenthawed bovine embryos. Theriogenology, 37, 23-37. Go to original source...
    55. White KL, Lindner GM, Anderson GB, BonDurant RH (1982): Survival after transfer of "sexed" mouse embryos exposed to H-Y antisera. Theriogenology, 18, 655-662. Go to original source...
    56. Williams TJ (1986): A technique for sexing mouse embryos by a visual colorimetric assay of the X-linked enzyme, glucose 6-phosphate dehydrogenase. Theriogenology, 25, 733-739. Go to original source... Go to PubMed...
    57. Yu W, Li S, Fang J, Sun X, Cui L, Fu J, Bai Y, Fang Y, Shangguan B (2007): Field studies on the effectiveness of the YCD embryo sexing technique in bovine. Reproduction, Fertility and Development, 19, 299-300. Go to original source...
    58. Zhang M, Lu KH, Seidel GE Jr. (2005): Blastocyst development of male and female bovine embryos produced by IVF with flow cytometrically sorted sperm. Reproduction, Fertility and Development, 17, 306-307. Go to original source...

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content