Birth of cloned calves produced with adult and fetal fibroblast

Authors

  • Marco Roberto Bourg de Mello Universidade Federal Rural do Rio de Janeiro, Instituto de Zootecnia, Departamento de Reprodução e Avaliação Animal, Seropédica, RJ
  • Heloísa Vasconcellos Amaral Caetano Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • Mariana Groke Marques Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • Marcella Pecora Milazzotto Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • Alessandra Corallo Nicácio Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • Camilla Mota Mendes Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • Viviane Purri Oliveira Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • Mayra Elena Ortiz D'Avila Assumpção Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP
  • José Antonio Visintin Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Reprodução Animal, São Paulo, SP

Keywords:

bovine, cloning, fibroblast, Nellore, nuclear transfer

Abstract

The present study examined the viability of bovine nuclear transferred embryos. Oocytes were matured in vitro for 17 hours and enucleated after aspiration of the first polar body and the metaphase plate. Fibroblasts from a 10-year-old Nellore cow and 45-day-old male fetus collected from slaughterhouse were used as nuclei donor. Enucleated oocytes were fused with adult and fetal fibroblasts with an electric stimulus. After electrical fusion, the couplets were incubated in TCM199 plus 10% FCS supplemented with cycloheximide and cytochalasin D for 1 hour and then cycloheximide alone for an additional 4 hours. The activated reconstructed embryos were co-cultured with granulosa cells in TCM 199 for 7–9 days. A total of 569 couplets were reconstructed from adult and 668 from fetal fibroblasts. After electrofusion, 181 (adult cells) and 212 (fetal cells) embryos became fused and 30 (16.6%) and 32 (15.1%) reached the blastocyst stage, respectively. After transferring 21 (adult cells) and 18 (fetal cells) blastocysts, pregnancy rates at day 90 were 19% (4) and 16.7% (3), respectively. There was no significant difference (P<0.05) between the pregnancy rates. Two pregnancies from adult cells aborted at the 4th and 5th months of gestation. One recipient that received an embryo from adult fibroblasts produced a healthy Nellore female calf weighting 40 kg. The other recipient that received an embryo from adult fibroblasts had hydrallantois and delivered a dead calf at day 290. The first pregnancy from fetal cells produced a healthy male calf, weighting 34kg (MELLO et al. 2003). These results indicated that adult and fetal Nellore fibroblasts could be used successfully as nuclei donor, with similar rates of in vitro and in vivo development.

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Published

2008-03-14

Issue

Section

ANIMAL REPRODUCTION

How to Cite

Birth of cloned calves produced with adult and fetal fibroblast. (2008). Bulletin of Animal Husbandry, 65(3), 217-223. http://bia.iz.sp.gov.br/index.php/bia/article/view/1129

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