Post-biopsy bovine embryo viability and whole genome amplification in preimplantation genetic diagnosis.
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.


METHODS
Randomized study.


METHODS
Molecular genetics and animal reproduction laboratories.


METHODS
Cow ovaries obtained from slaughterhouses.


METHODS
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.


METHODS
Subsequent rate of blastocyst development, embryo cell number, WGA efficiency, and sex determination.


RESULTS
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.


CONCLUSIONS
Biopsy of 8-cell to 16-cell bovine embryos did not affect their subsequent development. WGA was successful in removed blastomeres.
Polisseni
Juliana
J
Empresa Brasileira de Pesquisa Agropecuária, Embrapa Gado de Leite, Brazil. jupol@powermail.com.br
Sá
Wanderlei Ferreira de
WF
Guerra
Martha de Oliveira
Mde O
Machado
Marco Antônio
MA
Serapião
Raquel Varella
RV
Carvalho
Bruno Campos de
BC
Camargo
Luiz Sérgio de Almeida
LS
Peters
Vera Maria
VM
D000818:Animals; D001706:Biopsy; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D001757:Blastomeres / Q000166:cytology / Q000502:physiology*; D002417:Cattle; D046149:Embryo Culture Techniques / Q000662:veterinary; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D016678:Genome; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000662:veterinary*; D012732:Sex Determination Analysis / Q000662:veterinary