Post-biopsy bovine embryo viability and whole genome amplification in preimplantation genetic diagnosis
Received 4 July 2008; received in revised form 6 October 2008; accepted 17 October 2008. published online 25 December 2008.
Objective
To evaluate the effect of the biopsy of 8-cell to 16-cell bovine embryos on their subsequent development and the effect of whole genome amplification (WGA) on removed blastomeres.
Design
Randomized study.
Setting
Molecular genetics and animal reproduction laboratories.
Patient(s)
Cow ovaries obtained from slaughterhouses.
Intervention(s)
The ovaries were punctured, and the oocytes were matured and fertilized in vitro. On the fourth day after fertilization, 8-cell to 16-cell bovine embryos were biopsied, one quarter of each embryo being removed. The blastomeres were submitted to WGA followed by polymerase chain reaction (PCR). The embryos were returned to culture for evaluation of their development.
Main Outcome Measure(s)
Subsequent rate of blastocyst development, embryo cell number, WGA efficiency, and sex determination.
Result(s)
A total of 92 embryos were submitted to biopsy. The blastocyst production was 53.3%, with 44.9% of hatching rate. These results were similar to those of the control group (66.0% and 42.6%) of 103 embryos. Overall, no impact was detected on embryo quality in blastocyst cell number between the two groups. Removed blastomeres were submitted to WGA, resulting in 98.2% of efficiency. However, only 59% of the samples were sexed by PCR.
Conclusion(s)
Biopsy of 8-cell to 16-cell bovine embryos did not affect their subsequent development. WGA was successful in removed blastomeres.
aEmpresa Brasileira de Pesquisa Agropecuária, Embrapa Gado de Leite, Brazil
bCentro de Biologia da Reprodução, Universidade Federal de Juiz de Fora, Brazil
cEmpresa de Pesquisa Agropecuária de Minas Gerais, Epamig, Juiz de Fora, Minas Gerais, Brazil
Reprint requests: Juliana Polisseni, M.S., Rua Oscar Vidal 245/301, Centro Juiz de Fora, Minas Gerais, Brazil CEP:36016-290 (FAX: 55-32-3216-5092).
J.P. has nothing to disclose. W.F.de S. has nothing to disclose. M. de O.G. has nothing to disclose. M.A.M. has nothing to disclose. R.V.S. has nothing to disclose.
B.C. de C. has nothing to disclose. L.S. de A.C. has nothing to disclose. V.M.P. has nothing to disclose.
Supported by Fundação de Amparo à pesquisa do Estado de Minas Gerais (FAPEMIG), Belo Horizonte, Minas Gerais, Brazil Fapemig–Rede 2827/05); Centro Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília, Distrito Federal, Brazil (CNPq/480.205/2004-3).
ABSTRACT