SEPARATION OF X-BEARING BOVINE SPERM BY CENTRIFUGATION IN CONTINUOUS PERCOLL AND OPTIPREP DENSITY GRADIENT: EFFECT IN SPERM VIABILITY AND IN VITRO EMBRYO PRODUCTION

Authors

  • Max Vitória Resende Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brasil
  • Marcelo Barbosa Bezerra Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brasil
  • Felipe Perecin Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brasil
  • Adriana Oliveira Almeida Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brasil
  • Aline Costa Lucio Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brasil
  • Vera Fernanda Martins Hossepian de Lima Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brasil

Keywords:

bovino, espermatozóides X, centrifugação, produção in vitro de embriões, PCR

Abstract

The aim of this study was to separate X-bearing bovine sperm by continuous Percoll and OptiPrep density gradients and to validate the sexing of resultant in vitro produced embryos by Polimerase Chain Reaction (PCR). Frozen/thawed sperm was layered on density gradients which were previously prepared in polystyrene tubes, 24 h before procedures and maintained at 4 °C. The tubes were centrifuged at 500 x g for 15 min at 22 °C. Supernatants were gently aspirated and the sperm recovered from the bottom of the tubes. Viability and integrity of sperm were evaluated by Trypan Blue/Giemsa stain. Cleavage and blastocyst rates were determined by in vitro production of embryos and PCR was performed for identification of the embryos’ genetic sex. No damage in viability and acrossomal integrity and in cleavage and blastocyst rates was found in the Percoll and OptiPrep treatment compared to the non-centrifuged group (P>0.05). The percentage of female embryos in the Percoll and OptiPrep group was 63.0 and 47.6%, respectively. The female embryos in control group were 48.7%. A sexual deviation in the Percoll density gradient was achieved without reduction of sperm viability and in vitro production rates.

KEY WORDS: Bovine, centrifugation, in vitro production of embryos, PCR, X-bearing sperm.

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Published

2009-07-03

How to Cite

RESENDE, Max Vitória; BEZERRA, Marcelo Barbosa; PERECIN, Felipe; ALMEIDA, Adriana Oliveira; LUCIO, Aline Costa; LIMA, Vera Fernanda Martins Hossepian de. SEPARATION OF X-BEARING BOVINE SPERM BY CENTRIFUGATION IN CONTINUOUS PERCOLL AND OPTIPREP DENSITY GRADIENT: EFFECT IN SPERM VIABILITY AND IN VITRO EMBRYO PRODUCTION. Brazilian Animal Science/ Ciência Animal Brasileira, Goiânia, v. 10, n. 2, p. 581–587, 2009. Disponível em: https://revistas.ufg.br/vet/article/view/2345. Acesso em: 13 dec. 2025.

Issue

Vol. 10 No. 2 (2009)

Section

Veterinary Medicine

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Copyright (c) 2009 Brazilian Animal Science/ Ciência Animal Brasileira

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