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 Structure
All Issues
Special Issues
Research Fronts
Sample Issue
For Authors
General Information
Submit Article
Author Instructions
Open Access
Awards and Prizes
For Referees
Referee Guidelines
Review an 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 LinkedIn

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 >>   Contents Vol 25(6)

Development of bovine embryos derived from reproductive techniques

Míryan L. V. Alberto A, Flavio V. Meirelles B, Felipe Perecin B, Carlos E. Ambrósio B, Phelipe O. Favaron A, André L. R. Franciolli A, Andrea M. Mess A, José M. dos Santos C, Rose E. G. Rici A, Marcelo Bertolini D and Maria A. Miglino A E

A Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Av. Prof. Orlando Marques de Paiva, 87, Butantã, 05508-270, Sao Paulo, Brazil.
B Faculty of Animal Sciences and Food Engineering, University of Sao Paulo, Av. Duque de Caxias Norte, 225, Caixa Postal 23, 13635-900, Pirassununga, São Paulo, Brazil.
C Anhembi Morumbi University and School of Medicine from ABC, R. Dr. Almeida Lima, 1134, 03164-000, Sao Paulo, Brazil.
D University of Fortaleza, Av. Washington Soares, 1321, Edson Queiroz, 60.811-905, Fortaleza, Ceará, Brazil.
E Corresponding author. Email: miglino@usp.br

Reproduction, Fertility and Development 25(6) 907-917 http://dx.doi.org/10.1071/RD12092
Submitted: 24 February 2012  Accepted: 1 August 2012   Published: 30 August 2012

PDF (1.1 MB) $25
 Export Citation

Assisted reproduction techniques have improved agricultural breeding in the bovine. However, important development steps may differ from the situation in vivo and there is a high mortality rate during the first trimester of gestation. To better understand these events, we investigated the development of embryos and fetal membranes following fixed-time AI (FTAI), IVF and nuclear transfer (NT). The onset of yolk-sac development was not normal in cloned embryos. Later steps differed from conditions in vivo in all three groups; the yolk-sac was yellowish and juxtaposed with the amniotic membrane. Vascularisation of the chorioallantoic membrane was relatively late and low in NT gestations, but normal in the others. The overall development of the embryos was normal, as indicated by morphology and regression analysis of growth rate. However, NT conceptuses were significantly smaller, with the livers in some embryos occupying the abdominal cavity and others exhibiting heart abnormalities. In conclusion, the yolk-sac and the cardiovascular system seem to be vulnerable to morphogenetic alterations. Future studies will focus on gene expression and early vascularisation processes to investigate whether these changes may be responsible for the high incidence of intrauterine mortality, especially in clones.

Additional keywords: amnion, animal cloning, chorioallantoic membrane, fixed-time artificial insemination, heart development, IVF, lung development, nuclear transfer, yolk-sac.


Abu-Issa, R., Waldo, K., and Kirby, M. L. (2004). Heart fields: one, two or more? Dev. Biol. 272, 281–285.
CrossRef | CAS | PubMed |

Alves, D. F., Rauber, L. P., and Rubin, F. B. (2003). Desenvolvimento embrionário in vitro de oócitos bovinos mantidos em líquido folicular ou TCM-hepes. Braz. J. Vet. Res. Anim. Sci. 40, 279–286.
CrossRef |

Assis Neto, A. C., Pereira, F. T. V., Santos, T. C., Ambrósio, C. E., and Miglino, M. A. (2010). Morpho-physical recording of bovine conceptus (Bos indicus) and placenta from Days 20 to 70 of pregnancy. Reprod. Domest. Anim. 45, 760–772.
| CAS | PubMed |

Baruselli, P. S., Marques, M. O., Carvalho, N. A. T., Berber, R. C. A., Valentim, R., Carvalho Filho, A. F., and Costa Neto, W. P. (2003). Dinâmica folicular e taxa de prenhez em novilhas receptoras de embrião (Bos taurus indicus x Bos taurus taurus) tratadas com o protocolo ‘Ovsynch’ para inovulação em tempo fixo. Braz. J. Vet. Res. Anim. Sci. 40, 96–106.
CrossRef |

Batchelder, C. A., Bertolini, M., Manson, J. B., Moyer, A. L., Hoffert, K. A., Petkov, S. G., Famula, T. R., Angelos, J., George, L. W., and Anderson, G. B. (2007). Perinatal physiology in cloned and normal calves: physical and clinical characteristics. Cloning Stem Cells 9, 63–82.
CrossRef | CAS | PubMed |

Behboodi, E., Anederson, G. B., Bondurant, R. H., Cargill, S. L., Kreuscher, B. R., Medrano, J. F., and Murray, J. D. (1995). Birth of large calves that developed from in vitro-derived bovine embryos. Theriogenology 44, 227–232.
CrossRef | CAS | PubMed |

Bertolini, M., and Anderson, G. B. (2002). The placenta as a contributor to production of large calves. Theriogenology 57, 181–187.
CrossRef | CAS | PubMed |

Bertolini, M., Mason, J. B., Beam, S. W., Carneiro, G. F., Sween, M. L., Kominek, D. J., Moyer, A. L., Famula, T. R., Sainz, R. D., and Anderson, G. B. (2002). Morphology and morphometry of in vivo- and in vitro-produced bovine concepti from early pregnancy to term and association with high birth weights. Theriogenology 58, 973–994.
CrossRef | PubMed |

Bertolini, M., Moyer, A. L., Mason, J. B., Batchelder, C. A., Hoffert, K. A., Bertolini, L. R., Carneiro, G. F., Cargill, S. L., Famula, T. R., Calvert, C. C., Sainz, R. D., and Anderson, G. B. (2004). Evidence of increased substrate availability to in vitro-derived bovine fetuses and association with accelerated conceptus growth. Reproduction 128, 341–354.
CrossRef | CAS | PubMed |

Betteridge, K. (2003). A history of farm animal embryo transfer and some associated techniques. Anim. Reprod. Sci. 79, 203–244.
CrossRef | PubMed |

Block, J., Hansen, P. J., Loureiro, B., and Bonilla, L. (2011). Improving post-transfer survival of bovine embryos produced in vitro: actions of insulin-like growth factor-1, colony stimulating factor-2 and hyaluronan. Theriogenology 76, 1602–1609.
CrossRef | CAS | PubMed |

Brackett, B. G., Bousquet, D., Boice, M. L., Donawick, W. J., Evans, J. F., and Dressel, M. A. (1982). Normal development following in vitro fertilization in the cow. Biol. Reprod. 27, 147–158.
CrossRef | CAS | PubMed |

Bryden, M. M., Evans, H., and Binns, W. (1973). Embryology of the sheep. III. The respiratory system, mesenteries and celom in the fourteen to thirty-four day embryo. Anat. Rec. 175, 725–735.
CrossRef | CAS | PubMed |

Burri, P. H. (1984). Fetal and postnatal development of the lung. Annu. Rev. Physiol. 46, 617–628.
CrossRef | CAS | PubMed |

Campos, D. B., Papa, P. C., Marques, J. E., Garbelotti, F., Fátima, L. A., Artoni, L. P., Birgel, E. H., Meirelles, F. V., Buratini, J., Leiser, R., and Pfarrer, C. (2010). Somatic cell nuclear transfer is associated with altered expression of angiogenic factor systems in bovine placentomes at term. Genet. Mol. Res. 9, 309–323.
CrossRef | CAS | PubMed |

Chavatte-Palmer, P., Remy, D., Cordonnier, N., Richard, C., Issenman, H., and Laigre, P. (2004). Review: health status of cloned cattle at different ages. Cloning Stem Cells 6, 94–100.
CrossRef | CAS | PubMed |

Chavatte-Palmer, P., Sousa, N., Laigre, P., Camous, S., Ponter, A. A., Beckers, J. F., and Heyman, Y. (2006). Ultrasound fetal measurements and pregnancy associated glycoprotein secretion in early pregnancy in cattle recipients carryng somatic clones. Theriogenology 66, 829–840.
CrossRef | CAS | PubMed |

Chavatte-Palmer, P., Camousb, S., Jammes, H., Le Cleac’h, N., Guillomot, M., and Lee, R. S. F. (2012). Review: placental perturbations induce the developmental abnormalities often observed in bovine somatic cell nuclear transfer. Placenta 33, S99–S104.
| PubMed |

De Bem, T. H., Chiaratti, M. R., Rochetti, R., Bressan, F. F., Sangalli, J. R., Miranda, M. S., Pires, P. R., Schwartz, K. R., Sampaio, R. V., Fantinato-Neto, P., Pimentel, J. R., Perecin, F., Smith, L. C., Meirelles, F. V., Adona, P. R., and Leal, C. L. (2011). Viable calves produced by somatic cell nuclear transfer using meiotic-blocked oocytes. Cell Reprogram. 13, 419–429.
| CAS | PubMed |

De Sousa, P. A., King, T., Harkness, L., Young, L. E., Walker, S. K., and Wilmut, I. (2001). Evaluation of gestational deficiencies in cloned sheep fetuses and placentae. Biol. Reprod. 65, 23–30.
CrossRef | CAS | PubMed |

Drost, M. (2007). Complications during gestation in the cow. Theriogenology 68, 487–491.
CrossRef | CAS | PubMed |

Duplessis, H. (1970). ‘Embriologia: Cuadernos Practicos.’ (Toray-Massón: Barcelona.)

Edwards, J. L., Schrick, F. N., Mccracken, M. D., Van Amstel, S. R., Hopkins, F. M., Welborn, M. G., and Davies, C. J. (2003). Cloning adult farm animals: a review of the possibilities and problems associated with somatic cell nuclear transfer. Am. J. Reprod. Immunol. 50, 113–123.
CrossRef | CAS | PubMed |

Evans, H. E., and Sack, W. O. (1973). Prenatal development of domestic and laboratory mammals: growth curves, external features and selected references. Anat. Histol. Embryol. 2, 11–45.
CrossRef | CAS |

Farin, C. E., Farin, P. W., and Piedrahita, J. A. (2004). Development of fetuses in vitro-produced and cloned bovine embryos. J. Anim. Sci. 82, 53–62.

Farin, P. W., Piedrahita, J. A., and Farin, C. E. (2006). Errors in development of fetuses and placentas from in vitro-produced bovine embryos. Theriogenology 65, 178–191.
CrossRef | PubMed |

Gong, Z. J., Zhou, Y. Y., Xu, M., Cai, Q., Li, H., Yan, J. B., Wang, J., Zhang, H. J., Fan, S. Y., Yuan, Q., Huang, S. Z., and Zeng, F. (2012). Aberrant expression of imprinted genes and their regulatory network in cloned cattle. Theriogenology 78, 858–866.
CrossRef | CAS | PubMed |

Hansen, P. J., Block, J., Loureiro, B., Bonilla, L., and Hendricks, K. E. (2010). Effects of gamete source and culture conditions on the competence of in vitro-produced embryos for post-transfer survival in cattle. Reprod. Fertil. Dev. 22, 59–66.
CrossRef | PubMed |

Hill, J. R., Rousell, A. J., Cibelli, J. B., Edwards, J. F., Hooper, N. L., Miller, M. W., Thompson, J. A., Looney, C. R., Westhusin, M. E., Robl, J. M., and Stice, S. L. (1999). Clinical and pathlogic features of cloned transgenic calves and fetus (13 case studies). Theriogenology 51, 1451–1465.
CrossRef | CAS | PubMed |

Hill, J. R., Burghardt, R. C., Jones, K., Long, C. R., Looney, C. R., Shin, T., Spencer, T. E., Thompson, J. A., Winger, Q. A., and Westhusin, M. E. (2000). Evidence for placental abnormality as the major cause of mortality in first-trimester somatic cell cloned bovine fetuses. Biol. Reprod. 63, 1787–1794.
CrossRef | CAS | PubMed |

Hill, J. R., Edwards, J. F., Sawyer, N., Blackwell, C., and Cibelli, J. B. (2001). Placental anomalies in a viable cloned calf. Cloning 3, 83–88.
CrossRef | CAS | PubMed |

Hislop, A. A. (2002). Airway and blood vessel interaction during lung development. J. Anat. 201, 325–334.
CrossRef | PubMed |

Humblot, P., Le Bourhis, D., Fritz, S., Colleau, J. J., Gonzalez, C., Guyader Joly, C., Malafosse, A., Heyman, Y., Amigues, Y., Tissier, M., and Ponsart, C. (2010). Reproductive technologies and genomic selection in cattle. Vet. Med. Int. 24, 192787.

Hyttel, P., Sinowatz, F., and Vejlsted, M. (2010). ‘Essentials of Domestic Animal Embryology.’ (Saunders Elsevier: Toronto.)

Ioannides, A. S., Hederson, D. J., Spitz, L., and Copp, A. J. (2003). Role of sonic hedgehog in the development of the trachea and oesophagus. J. Pediatr. Surg. 38, 29–36.
CrossRef | PubMed |

Leidenfrost, S., Boelhauve, M., Reichenbach, M., Güngör, T., Reichenbach, H. D., Sinowatz, F., Wolf, E., and Habermann, F. A. (2011). Cell arrest and cell death in mammalian preimplantation development: lessons from the bovine model. PLoS One 6, e22121.
CrossRef | CAS | PubMed |

Lonergan, P., Evans, A. C. O., Boland, E., Rizos, D., Fair, T., Duffy, P., Sung, L. Y., Du, F., Chaubal, S., Xu, J., Yang, X., and Tian, X. C. (2007). Pregnancy and fetal characteristics after transfer of vitrified in vivo and cloned bovine embryos. Theriogenology 68, 1128–1137.
CrossRef | CAS | PubMed |

Matsuzaki, M., and Shiga, K. (2002). Endocrine characteristics of cloned calves. Cloning Stem Cells 4, 261–267.
CrossRef | CAS | PubMed |

Meirelles, F. V., Birgel, E. H., Perecin, F., Bertolini, M., Traldi, A. S., Pimentel, J. R., Komninou, E. R., Sangalli, J. R., Neto, P. F., Nunes, M. T., Pogliani, F. C., Meirelles, F. D., Kubrusly, F. S., Vannucchi, C. I., and Silva, L. C. (2010). Delivery of cloned offspring: experience in Zebu cattle (Bos indicus). Reprod. Fertil. Dev. 22, 88–97.
CrossRef | PubMed |

Mello, M. R. B., Caetano, H. V. A., Marques, M. G., Padilha, M. S., Garcia, J. F., Milazzotto, M. P., Assumpção, M. E. O. A., Lima, A. S., Nicácio, A. C., Mendes, C. M., Oliveira, V. P., and Visintin, J. A. (2003). Production of a cloned calf from a fetal fibroblast cell line. Braz. J. Med. Biol. Res. 36, 1485–1489.
CrossRef | CAS |

Miglino, M. A., Pereira, F. T. V., Visintin, J. A., Garcia, J. M., Meirelles, F. V., Rumpf, R., Ambrósio, C. E., Papa, P. C., Santos, T. C., Carvalho, A. F., Leiser, R., and Carter, A. M. (2007). Placentation in cloned cattle: structure and microvascular architecture. Theriogenology 68, 604–617.
CrossRef | CAS | PubMed |

Moore, K. L. and Persaud, T. V. N. (2004). ‘Embriologia Clínica.’ (Elsevier: Rio de Janeiro.)

Moore, K. L., Persaud, T. V., and Shiota, K. (2002). ‘Atlas Colorido de Embriologia Clínica.’ (Guanabara Koogan: Rio de Janeiro.)

Mundim, T. C., Ramos, A. F., Sartori, R., Dode, M. A., Melo, E. O., Gomes, L. F., Rumpf, R., and Franco, M. M. (2009). Changes in gene expression profiles of bovine embryos produced in vitro, by natural ovulation, or hormonal superstimulation. Genet. Mol. Res. 8, 1398–1407.
CrossRef | CAS | PubMed |

Noden, D. M., and De Lahunta, A. (1990). ‘Embriología de los Animales Domésticos: Mecanismo de Desarrollo y Malformaciones.’ (Acribia: Zaragoza.)

Ogura, A., Inoue, K., Ogonuki, N., Lee, J., Kohda, T., and Ishino, F. (2002). Phenotypic effects of somatic cell cloning in the mouse. Cloning Stem Cells 4, 397–405.
CrossRef | CAS | PubMed |

Reichenbach, H-D., Arnold, G. J., Bauersachs, S., and Beier, H. M. (2003). Morte embrionária na TE: redução através da melhor compreensão dos mecanismos da comunicação materno-embrionária? O Embrião 17, 3–6.

Ripamonte, P., Mesquita, L. G., Cortezzi, S. S., de Carvalho Balieiro, J. C., Fonseca Merighe, G. K., Watanabe, Y. F., Caetano, A. R., and Meirelles, F. V. (2011). Differential gene expression and developmental competence in in vitro produced bovine embryos. Zygote 15, 1–10.

Rodríguez-Alvarez, L., Sharbati, J., Sharbati, S., Cox, J. F., Einspanier, R., and Castro, F. O. (2010). Differential gene expression in bovine elongated (Day 17) embryos produced by somatic cell nucleus transfer and in vitro fertilization. Theriogenology 74, 45–59.
CrossRef | PubMed |

Rodriguez-Martinez, H. (2012). Assisted reproductive techniques for cattle breeding breeding in developing countries: a critical appraisal of their value and limitations. Reprod. Domest. Anim. 47, 21–26.
CrossRef | PubMed |

Ross, P. J., and Cibelli, J. B. (2010). Bovine somatic cell nuclear transfer. Methods Mol. Biol. 636, 155–177.
CrossRef | PubMed |

Rüsse, I., Sinowatz, F., Richter, L., Lehmann, M., and Schallenberger, E. (1992). The development of the yolk sac in ruminants (sheep and cattle). Anat. Histol. Embryol. 21, 324–347.
| PubMed |

Saadeldin, I. M., Kim, B., Lee, B., and Jang, G. (2011). Effect of different culture media on the temporal gene expression in the bovine developing embryos. Theriogenology 75, 995–1004.
CrossRef | CAS | PubMed |

Sakai, R. R., Tamashiro, L. K., Yamazaki, Y., and Yanagimachi, R. (2005). Cloning and assisted reproductive techniques: influence on early development and adult phenotype. Birth Defects Res. C Embryo Today 75, 151–162.
CrossRef | CAS | PubMed |

Sangalli, J. R., De Bem, T. H., Perecin, F., Chiaratti, M. R., de Oliveira, L. J., de Araújo, R. R., Valim Pimentel, J. R., Smith, L. C., and Meirelles, F. V. (2012). Treatment of nuclear-donor cells or cloned zygotes with chromatin-modifying agents increases histone acetylation but does not improve full-term development of cloned cattle. Cell Reprogram. 14, 235–247.
| CAS | PubMed |

Stringfellow, D. A. and Seidel, S. M. (1998). ‘Manual da Sociedade Internacional de Transferência de Embriões: Um Guia de Procedimentos e Informações Gerais Para uso em Tecnologia de Transferência de Embriões Enfatizando Procedimentos Sanitários.’ (Savoy: Illinois.)

Vitums, A. (1981). The embryonic development of the equine heart. Anat. Histol. Embryol. 10, 193–211.
CrossRef | CAS | PubMed |

Watanabe, Y. F. (1998). A fecundação in vitro e a reação acrossomal como critério de seleção para fertilidade em touros da raça Nelore [In vitro fertilization and acrosome reaction as selection criteria for fertility in Nellore cows]. Ph.D. Thesis, University of São Paulo, Brazil.

Wells, D., Misica, P., and Tervit, H. (1999). Production of cloned calves following nuclear transfer with cultured adult mural granulose cells. Biol. Reprod. 60, 996–1005.
CrossRef | CAS | PubMed |

Wells, D. N., Forsyth, J. T., McMillan, V., and Oback, B. (2004). The health of somatic cell cloned cattle and their offspring. Cloning Stem Cells 6, 101–110.
CrossRef | CAS | PubMed |

Winters, L. M., Green, W. W., and Constock, R. E. (1942). ‘Prenatal Development of the Bovine.’ Volume 151 of Technical Bulletin (University of Minnesota, Agricultural Experiment Station, St Paul.)

Yamazaki, W., Ferreira, C. R., Méo, S. C., Leal, C. L., Meirelles, F. V., and Garcia, J. M. (2005). Use of strontium in the activation of bovine oocytes reconstructed by somatic cell nuclear transfer. Zygote 13, 295–302.
CrossRef | CAS | PubMed |

Young, L. E., Sinclair, K. D., and Wilmut, I. (1998). Large offspring syndrome in cattle and sheep. J. Reprod. Fertil. 3, 155–163.
| CAS |

Subscriber Login

Legal & Privacy | Contact Us | Help


© CSIRO 1996-2015