In Vitro Culture of Embryos from LOPU-Derived Goat Oocytes

  • Joanna Maria G. Souza-FabjanEmail author
  • Ribrio Ivan T. P. Batista
  • Vicente José F. Freitas
  • Pascal Mermillod
Part of the Methods in Molecular Biology book series (MIMB, volume 2006)


A high oocyte quality is the prerequisite for in vitro embryo production. Goat cumulus–oocyte complexes (COC) are mainly collected from slaughterhouse ovaries or by laparoscopic ovum pickup (LOPU) from live animals. Several features can influence the availability of good quality oocytes recovered by the LOPU technique. Interestingly, slaughterhouse and LOPU oocytes have different in vitro maturation kinetics and requirements, and thus, the IVP system must be adapted regarding the oocyte origin. Overall, the use of undefined media in the different steps makes interpretation of results more difficult, hampers their reproducibility, and introduces a sanitary risk. Thus, there is an effort worldwide to use simpler conditions for goat IVP. Although the success of IVP rates is relatively high, in vitro embryos differ from in vivo-derived ones in many aspects, resulting in lower viability. Therefore, strategies to improve in vitro embryo quality are crucial, such as the use of oviductal epithelium cells for coculture. Here we describe the main steps and culture media which can be utilized to produce embryos in vitro from LOPU or slaughterhouse oocytes in goats.

Key words

Caprine IVF IVP Goat COC Laboratory protocol OPU 



The authors wish to thank CAPES-COFECUB bilateral framework for financial support of collaboration between the State University of Ceará and INRA. J.M.G. Souza-Fabjan and R.I.T.P. Batista conducted their postdoctoral studies under the auspices of CAPES (Brasília, Brazil), J.M.G. Souza-Fabjan is a fellow of FAPERJ, and V.J.F. Freitas is a fellow of CNPq.


  1. 1.
    Duszewska AM, Trzeciak P, Compa A et al (2010) Selected issues concerning biotechnology of farm animals breeding—a review. Anim Sci Pap Rep 28:295–306Google Scholar
  2. 2.
    Souza-Fabjan JMG, Panneau B, Duffard N et al (2014a) In vitro production of small ruminant embryos: late improvements and further research. Theriogenology 81:1149–1162CrossRefGoogle Scholar
  3. 3.
    Paramio MT, Izquierdo D (2016) Recent advances in in vitro embryo production in small ruminants. Theriogenology 86:152–159CrossRefGoogle Scholar
  4. 4.
    Baldassarre H, Karatzas CN (2004) Advanced assisted reproduction technologies (ART) in goats. Anim Reprod Sci 82–83:255–266CrossRefGoogle Scholar
  5. 5.
    Sanchez DJD, Melo CHS, Souza-Fabjan JMG et al (2014) Repeated hormonal treatment and laparoscopic ovum pick-up followed by in vitro embryo production in goats raised in the tropics. Livest Sci 165:217–222CrossRefGoogle Scholar
  6. 6.
    Teixeira PPM, Padilha LC, Oiveira MEF et al (2011) Laparoscopic ovum collection in sheep: gross and microscopic evaluation of the ovary and influence on oocyte. Anim Reprod Sci 127:169–175CrossRefGoogle Scholar
  7. 7.
    Souza-Fabjan JMG, Locatelli Y, Freitas VJF et al (2014b) Laparoscopic ovum pick up (LOPU) in goats: from hormonal treatment to oocyte possible destinations. R Bras Ci Vet 21:3–11CrossRefGoogle Scholar
  8. 8.
    Gibbons A, Pereyra Bonnet F, Cueto MI et al (2008) Recovery of sheep and goat oocytes by laparoscopy. Acta Sci Vet 36:223–230Google Scholar
  9. 9.
    Katska-Ksiazkiewicz L, Rynska B, Gajda B et al (2004) Effect of donor stimulation, frozen semen and heparin treatment on the efficiency of in vitro embryo production in goats. Theriogenology 62:576–586CrossRefGoogle Scholar
  10. 10.
    Souza-Fabjan JMG, Locatelli Y, Duffard N et al (2016) Intrinsic quality of goat oocytes already found denuded at collection for in vitro embryo production. Theriogenology 86:1989–1998CrossRefGoogle Scholar
  11. 11.
    Souza JMG, Duffard N, Bertoldo MJ et al (2013) Influence of heparin or the presence of cumulus cells during fertilization on the in vitro production of goat embryos. Anim Reprod Sci 138:82–89CrossRefGoogle Scholar
  12. 12.
    Souza-Fabjan JMG, Locatelli Y, Duffard N et al (2014c) In vitro embryo production in goats: Slaughterhouse and laparoscopic ovum pick up–derived oocytes have different kinetics and requirements regarding maturation media. Theriogenology 81:1021–1031CrossRefGoogle Scholar
  13. 13.
    Souza-Fabjan JMG, Pereira AF, Melo CHS et al (2013) Assessment of the reproductive parameters, laparoscopic oocyte recovery and the first embryos produced in vitro from endangered Canindé goats (Capra hircus). Reprod Biol 13:325–332CrossRefGoogle Scholar
  14. 14.
    Olivares CCS, Fonseca JF, Camargo LSA et al (2015) Comparison of different methods of goat sperm selection and capacitation for optimization of assisted reproductive technologies. Small Rumin Res 127:44–49CrossRefGoogle Scholar
  15. 15.
    Palomo MJ, Izquierdo D, Mogas T et al (1999) Effect of semen preparation on IVF of prepubertal goat oocytes. Theriogenology 51:927–940CrossRefGoogle Scholar
  16. 16.
    De Graaf SP, Beilby KH, Underwood SL (2009) Sperm sexing in sheep and cattle: the exception and the rule. Theriogenology 71:89–97CrossRefGoogle Scholar
  17. 17.
    Pontes JHF, Silva KCF, Basso AC (2010) Large-scale in vitro embryo production and pregnancy rates from Bos taurus, Bos indicus, and indicus-taurus dairy cows using sexed sperm. Theriogenology 74:1349–1355CrossRefGoogle Scholar
  18. 18.
    Cox JF, Alfaro V (2007) In vitro fertilization and development of OPU derived goat and sheep oocytes. Reprod Domest Anim 42:83–87CrossRefGoogle Scholar
  19. 19.
    Parrish JJ, Susko-Parrish J, Winer MA et al (1988) Capacitation of bovine sperm by heparin. Biol Reprod 38:1171–1180CrossRefGoogle Scholar
  20. 20.
    Malik RK, Lohan IS, Dhanda OP et al (1997) Test for the acrosomal reaction of goat spermatozoa treated with heparin. Small Rumin Res 26:163–166CrossRefGoogle Scholar
  21. 21.
    Paramio MT, Izquierdo D (2014) Current status of in vitro embryo production in sheep and goats. Reprod Domest Anim 49(Suppl. 4):37–48CrossRefGoogle Scholar
  22. 22.
    Wani AR, Khan MZ, Sofi KA et al (2012) Effect of cysteamine and epidermal growth factor (EGF) supplementation in maturation medium on in vitro maturation, fertilization and culturing of embryos in sheep. Small Rumin Res 106:160–164CrossRefGoogle Scholar
  23. 23.
    Katska-Ksiazkiewicz L, Opiela J, Rynska B (2007) Effects of oocyte quality, sêmen donor and embryo co-culture system on the efficiency of blastocyst production in goats. Theriogenology 68:736–744CrossRefGoogle Scholar
  24. 24.
    Beilby KH, de Graaf SP, Evans G et al (2011) Quantitative mRNA expression in ovine blastocysts produced from X- and Y chromosome bearing sperm, both in vitro and in vivo. Theriogenology 76:471–481CrossRefGoogle Scholar
  25. 25.
    Tervit HR, Whittingham DG, Rowson LE (1972) Successful culture in vitro of sheep and cattle ova. J Reprod Fertil 30:493–497CrossRefGoogle Scholar
  26. 26.
    Catalá MG, Izquierdo D, Rodríguez-Prado M (2012) Effect of oocyte quality on blastocyst development after in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in a sheep model. Fertil Steril 97:1004–1008CrossRefGoogle Scholar
  27. 27.
    Hammami S, Morató R, Romaguera R et al (2013) Developmental competence and embryo quality of small oocytes from pre-pubertal goats cultured in IVM medium supplemented with low level of hormones, insulin–transferrin–selenium and ascorbic acid. Reprod Domest Anim 48:339–344CrossRefGoogle Scholar
  28. 28.
    Wang Z, Lin P, Yu S (2013) Effects of ghrelin on developmental competence and gene expression of in vitro fertilized ovine embryos. Theriogenology 79:695–701CrossRefGoogle Scholar
  29. 29.
    Thompson JG, Partridge RJ, Houghton FD (1996) Oxygen uptake and carbohydrate metabolism by in vitro derived bovine embryos. J Reprod Fertil 106:299–306CrossRefGoogle Scholar
  30. 30.
    Gandolfi F, Moor RM (1987) Stimulation of early embryonic development in the sheep by co-culture with oviduct epithelial cells. J Reprod Fertil 81:23–28CrossRefGoogle Scholar
  31. 31.
    Bavister BD (1995) Culture of preimplantation embryos: facts and artifacts. Hum Reprod Update 1:91–148CrossRefGoogle Scholar
  32. 32.
    Rodríguez-Dorta N, Cognié Y, González F et al (2007) Effect of coculture with oviduct epithelial cells on viability after transfer of vitrified in vitro produced goat embryos. Theriogenology 68(6):908–913CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Joanna Maria G. Souza-Fabjan
    • 1
    Email author
  • Ribrio Ivan T. P. Batista
    • 1
  • Vicente José F. Freitas
    • 2
  • Pascal Mermillod
    • 3
  1. 1.Faculdade de Veterinária, Setor de Reprodução AnimalUniversidade Federal FluminenseNiteroiBrazil
  2. 2.Laboratório de Fisiologia e Controle da ReproduçãoUniversidade Estadual do CearáFortalezaBrazil
  3. 3.Institut National de Recherche Agronomique (INRA), UMR7247Physiologie de la Reproduction et des ComportementsNouzillyFrance

Personalised recommendations