CSIRO Publishing blank image blank image blank image blank imageBooksblank image blank image blank image blank imageJournalsblank image blank image blank image blank imageAbout Usblank image blank image blank image blank imageShopping Cartblank image blank image blank image You are here: Journals > Reproduction, Fertility and Development   
Reproduction, Fertility and Development
Journal Banner
  Vertebrate Reproductive Science & Technology
blank image Search
blank image blank image
blank image
  Advanced Search

Journal Home
About the Journal
Editorial Board
Online Early
Current Issue
Just Accepted
All Issues
Special Issues
Research Fronts
Sample Issue
For Authors
General Information
Instructions to Authors
Submit Article
Open Access
For Referees
Referee Guidelines
Review Article
Annual Referee Index
For Subscribers
Subscription Prices
Customer Service
Print Publication Dates

blue arrow e-Alerts
blank image
Subscribe to our email Early Alert or RSS feeds for the latest journal papers.

red arrow Connect with us
blank image
facebook twitter youtube

red arrow Connect with SRB
blank image
facebook TwitterIcon

Affiliated Societies

RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.


Article << Previous     |     Next >>        Online Early    

Selection of pre- versus postpubertal pig oocytes for parthenogenetic activation and somatic cell nuclear transfer

H. S. Pedersen A D , Y. Liu A , R. Li A , S. Purup A , P. Løvendahl B , P. Holm C , P. Hyttel C and H. Callesen A

A Department of Animal Science, Aarhus University, DK-8830 Tjele, Denmark.
B Department of Molecular Biology and Genetics, Aarhus University, DK-8830 Tjele, Denmark.
C Department of Veterinary Clinical and Animal Science, University of Copenhagen, DK-1870 Frederiksberg C, Denmark.
D Corresponding author. Email: hanne.skovsgaard@agrsci.dk

Reproduction, Fertility and Development - http://dx.doi.org/10.1071/RD13283
Submitted: 2 September 2013  Accepted: 10 January 2014   Published online: 24 February 2014

PDF (539 KB) $25
 Export Citation

Pig oocytes have been used increasingly for in vitro production techniques in recent years. The slaughterhouse-derived oocytes that are often used are mostly of prepubertal origin. The aims of the present study were to compare the developmental competence between pre- and postpubertal pig oocytes, and to develop a simple and practical method for the selection of prepubertal pig oocytes for parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) based on oocyte morphology after IVM and oocyte inside zona pellucida (ZP) diameter (‘small’ ≤110 µm; ‘medium’ >110 µm; ‘large’ ≥120 µm). Meiotic competence and blastocyst rates after PA and SCNT of prepubertal oocytes increased with oocyte size, with the large prepubertal oocytes reaching a level similar to postpubertal oocytes after SCNT. Blastocyst cell number was not related to oocyte inside ZP diameter and oocyte donor to the same extent as blastocyst rate. Very low blastocyst rates were obtained after PA of morphologically bad pre- and postpubertal oocytes. In conclusion, measurement of inside ZP diameter combined with morphological selection is useful to remove incompetent oocytes. Further studies are needed to clarify the relative importance of cytoplasmic volume and stage in oocyte growth phase.

Additional keywords: developmental competence, oocyte diameter, oocyte morphology.


Al-Mashhadi, R. H., Sorensen, C. B., Kragh, P. M., Christoffersen, C., Mortensen, M. B., Tolbod, L. P., Thim, T., Du, Y. T., Li, J., Liu, Y., Moldt, B., Schmidt, M., Vajta, G., Larsen, T., Purup, S., Bolund, L., Nielsen, L. B., Callesen, H., Falk, E., Mikkelsen, J. G., and Bentzon, J. F. (2013). Familial hypercholesterolemia and atherosclerosis in cloned minipigs created by DNA transposition of a human PCSK9 gain-of-function mutant. Sci. Transl. Med. 5, 166ra1.
CrossRef | PubMed |

Bagg, M. A., Vassena, R., Papasso-Brambilla, E., Grupen, C. G., Armstrong, D. T., and Gandolfi, F. (2004). Changes in ovarian, follicular, and oocyte morphology immediately after the onset of puberty are not accompanied by an increase in oocyte developmental competence in the pig. Theriogenology 62, 1003–1011.
CrossRef | CAS | PubMed |

Bagg, M. A., Nottle, M. B., Armstrong, D. T., and Grupen, C. G. (2007). Relationship between follicle size and oocyte developmental competence in prepubertal and adult pigs. Reprod. Fertil. Dev. 19, 797–803.
CrossRef | CAS | PubMed |

Ebner, T., Moser, M., Sommergruber, M., and Tews, G. (2003). Selection based on morphological assessment of oocytes and embryos at different stages of preimplantation development: a review. Hum. Reprod. Update 9, 251–262.
CrossRef | CAS | PubMed |

Fair, T., Hulshof, S. C. J., Hyttel, P., Greve, T., and Boland, M. (1997). Oocyte ultrastructure in bovine primordial to early tertiary follicles. Anat. Embryol. 195, 327–336.
CrossRef | CAS | PubMed |

Grupen, C. G., McIlfatrick, S. M., Ashman, R. J., Boquest, A. C., Armstrong, D. T., and Nottle, M. B. (2003). Relationship between donor animal age, follicular fluid steroid content and oocyte developmental competence in the pig. Reprod. Fertil. Dev. 15, 81–87.
CrossRef | PubMed |

Gupta, M. K., Uhm, S. J., and Lee, H. T. (2008). Sexual maturity and reproductive phase of oocyte donor influence the developmental ability and apoptosis of cloned and parthenogenetic porcine embryos. Anim. Reprod. Sci. 108, 107–121.
CrossRef | PubMed |

Hiraga, K., Hoshino, Y., Tanemura, K., and Sato, E. (2013). Selection of in vitro-matured porcine oocytes based on localization patterns of lipid droplets to evaluate developmental competence. J. Reprod. Dev. 59, 405–408.
CrossRef | CAS | PubMed |

Hunter, M. G. (2000). Oocyte maturation and ovum quality in pigs. Rev. Reprod. 5, 122–130.
CrossRef | CAS | PubMed |

Hyttel, P., Xu, K. P., Smith, S., and Greve, T. (1986). Ultrastructure of in-vitro oocyte maturation in cattle. J. Reprod. Fertil. 78, 615–625.
CrossRef | CAS | PubMed |

Ikeda, K., and Takahashi, Y. (2003). Comparison of maturational and developmental parameters of oocytes recovered from prepubertal and adult pigs. Reprod. Fertil. Dev. 15, 215–221.
CrossRef | CAS | PubMed |

Jakobsen, J. E., Li, J., Kragh, P. M., Moldt, B., Lin, L., Liu, Y., Schmidt, M., Winther, K. D., Schyth, B. D., Holm, I. E., Vajta, G., Bolund, L., Callesen, H., Jorgensen, A. L., Nielsen, A. L., and Mikkelsen, J. G. (2011). Pig transgenesis by Sleeping Beauty DNA transposition. Transgenic Res. 20, 533–545.
CrossRef | CAS | PubMed |

Kanayama, N., Miyano, T., and Lee, J. (2002). Acquisition of meiotic competence in growing pig oocytes correlates with their ability to activate Cdc2 kinase and MAP kinase. Zygote 10, 261–270.
CrossRef | CAS | PubMed |

Kim, J., You, J., Hyun, S. H., Lee, G., Lim, J., and Lee, E. (2010). Developmental competence of morphologically poor oocytes in relation to follicular size and oocyte diameter in the pig. Mol. Reprod. Dev. 77, 330–339.
| CAS | PubMed |

Kragh, P. M., Vajta, G., Corydon, T. J., Purup, S., Bolund, L., and Callesen, H. (2004). Production of transgenic porcine blastocysts by hand-made cloning. Reprod. Fertil. Dev. 16, 315–318.
CrossRef | CAS | PubMed |

Li, J., Svarcova, O., Villemoes, K., Kragh, P. M., Schmidt, M., Bogh, I. B., Zhang, Y., Du, Y., Lin, L., Purup, S., Xue, Q., Bolund, L., Yang, H., Maddox-Hyttel, P., and Vajta, G. (2008). High in vitro development after somatic cell nuclear transfer and trichostatin A treatment of reconstructed porcine embryos. Theriogenology 70, 800–808.
CrossRef | CAS | PubMed |

Li, J., Pedersen, H. S., Li, R., Adamsen, J., Liu, Y., Schmidt, M., Purup, S., and Callesen, H. (2013). Developmental potential of pig embryos reconstructed by use of sow versus pre-pubertal gilt oocytes after somatic cell nuclear transfer. Zygote 18, 1–10.
CrossRef |

Liu, Y., Ostrup, O., Li, J., Vajta, G., Kragh, P. M., Purup, S., and Callesen, H. (2011). Cell colony formation induced by Xenopus egg extract as a marker for improvement of cloned blastocyst formation in the pig. Cell. Reprogram. 13, 521–526.
| CAS | PubMed |

Lucas, X., Martinez, E. A., Roca, J., Vazquez, J. M., Gil, M. A., Pastor, L. M., and Alabart, J. L. (2002). Relationship between antral follicle size, oocyte diameters and nuclear maturation of immature oocytes in pigs. Theriogenology 58, 871–885.
CrossRef |

Luo, Y., Li, J., Liu, Y., Lin, L., Du, Y., Li, S., Yang, H., Vajta, G., Callesen, H., Bolund, L., and Sørensen, C. (2011). High efficiency of BRCA1 knockout using rAAV-mediated gene targeting: developing a pig model for breast cancer. Transgenic Res. 20, 975–988.
CrossRef | CAS | PubMed |

Marchal, R., Feugang, J. M., Perreau, C., Venturi, E., Terqui, M., and Mermillod, P. (2001). Meiotic and developmental competence of prepubertal and adult swine oocytes. Theriogenology 56, 17–29.
CrossRef | CAS | PubMed |

Marchal, R., Vigneron, C., Perreau, C., Bali-Papp, A., and Mermillod, P. (2002). Effect of follicular size on meiotic and developmental competence of porcine oocytes. Theriogenology 57, 1523–1532.
CrossRef | CAS | PubMed |

Mikkelsen, A. L., and Lindenberg, S. (2001). Morphology of in-vitro matured oocytes: impact on fertility potential and embryo quality. Hum. Reprod. 16, 1714–1718.
CrossRef | CAS | PubMed |

Motlik, J., Crozet, N., and Fulka, J. (1984). Meiotic competence in vitro of pig oocytes isolated from early antral follicles. J. Reprod. Fertil. 72, 323–328.
CrossRef | CAS | PubMed |

O’Brien, J. K., Dwarte, D., Ryan, J. P., Maxwell, W. M. C., and Evans, G. (2000). Comparison of in vitro maturation, in vitro fertilization, metabolism and ultrastructure of oocytes from prepubertal and adult pigs. Reprod. Domest. Anim. 35, 101–107.

Pedersen, H. S., Li, R., Liu, Y., Løvendahl, P., Holm, P., Hyttel, P., and Callesen, H. (2013). Development capacity of pre- and postpubertal pig oocytes evaluated by somatic cell nuclear transfer and parthenogenetic activation. Reprod. Fertil. Dev. 25, 241–242.
CrossRef |

Peura, T. T., Lewis, I. M., and Trounson, A. O. (1998). The effect of recipient oocyte volume on nuclear transfer in cattle. Mol. Reprod. Dev. 50, 185–191.
CrossRef | CAS | PubMed |

Rath, D., Freitag, K., Niemann, H., and Sieg, B. (1998). Continuous control of porcine embryo development in vitro. Theriogenology 49, 215.
CrossRef |

Schmidt, M., Winter, K. D., Dantzer, V., Li, J., Kragh, P. M., Du, Y., Lin, L., Liu, Y., Vajta, G., Sangild, P. T., Callesen, H., and Agerholm, J. S. (2011). Maternal endometrial oedema may increase perinatal mortality of cloned and transgenic piglets. Reprod. Fertil. Dev. 23, 645–653.
CrossRef | CAS | PubMed |

Sherrer, E. S., Rathbun, T. J., and Davis, D. L. (2004). Fertilization and blastocyst development in oocytes obtained from prepubertal and adult pigs. J. Anim. Sci. 82, 102–108.
| CAS | PubMed |

Sun, Q. Y., Wu, G. M., Lai, L., Park, K. W., Cabot, R., Cheong, H. T., Day, B. N., Prather, R. S., and Schatten, H. (2001). Translocation of active mitochondria during pig oocyte maturation, fertilization and early embryo development in vitro. Reproduction 122, 155–163.
CrossRef | CAS | PubMed |

Vajta, G., Peura, T. T., Holm, P., Páldi, A., Greve, T., Trounson, A. O., and Callesen, H. (2000). New method for culture of zona-included or zona-free embryos: The well of the well (WOW) system. Mol. Reprod. Dev. 55, 256–264.
CrossRef | CAS | PubMed |

Vajta, G., Lewis, I. M., Trounson, A. O., Purup, S., Maddox-Hyttel, P., Schmidt, M., Pedersen, H. G., Greve, T., and Callesen, H. (2003). Handmade somatic cell cloning in cattle: analysis of factors contributing to high efficiency in vitro. Biol. Reprod. 68, 571–578.
CrossRef | CAS | PubMed |

Westhusin, M. E., Collas, P., Marek, D., Sullivan, E., Stepp, P., Pryor, J., and Barnes, F. (1996). Reducing the amount of cytoplasm available for early embryonic development decreases the quality but not quantity of embryos produced by in vitro fertilization and nuclear transplantation. Theriogenology 46, 243–252.
CrossRef | CAS | PubMed |

Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I. M. K., and Iwamura, S. (2002). Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol. Reprod. 66, 112–119.
CrossRef | CAS | PubMed |

Zakhartchenko, V., Stojkovic, M., Brem, G., and Wolf, E. (1997). Karyoplast–cytoplast volume ratio in bovine nuclear transfer embryos: effect on developmental potential. Mol. Reprod. Dev. 48, 332–338.
CrossRef | CAS | PubMed |

Subscriber Login

Legal & Privacy | Contact Us | Help


© CSIRO 1996-2014