Research Article

Effect of culture media on porcine embryos produced by in vitro fertilization or parthenogenetic activation after oocyte maturation with cycloheximide

Mariana Groke Marques^a1 c1, Anibal Ballarotti Nascimento^a2, Renato Pereira da Costa Gerger^a2, José Sergio de Arruda Gonçalves^a2, Ana Rita de Sousa Coutinho^a2, Renata Simões^a2, Mayra Elena Ortiz D'Avila Assumpção^a2 and José Antônio Visintin^a2

^a1 Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva no. 87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.

^a2 Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.

Summary

This study evaluated the effects of reversible meiotic inhibition and different culture media (PZM3 or NCSU23) on production of porcine embryos by either in vitro fertilization (IVF) or parthenogenetic activation (PA). Oocytes from abattoir-derived ovaries were allocated into two groups for maturation: CHX (5 μg/ml cycloheximide for 10 h) or Control (no CHX). The percentage of metaphase II (MII) oocytes was determined at 36, 40 or 44 h of in vitro maturation. For IVF and PA, denuded oocytes were fertilized with purified sperm for 6 h or activated by electric stimuli. Zygotes were then subdivided into two culture groups: NCSU23 or PZM3. No effect of treatment with CHX and culture media was observed on cleavage (D3) and blastocyst (D7) rates in IVF and PA groups. There are no differences of quality or development rates between IVF-derived embryos cultured in NCSU23 or PZM3. However, we observed high quality PA embryos in PZM3 compared with NCSU23. Maturation arrest with CHX decreased the average blastocyst cell number in IVF while it was increased in PA embryos. As older oocytes are more effectively activated, CHX– blocked oocytes reached the mature stage faster than the control group. In conclusion, the CHX treatment for 10 h, followed by oocyte maturation for 40 h, is an efficient protocol to produce high quality parthenote embryos, especially when they are cultured in PZM3. However, this protocol is not satisfactory for IVF embryos production. In this case, a shorter maturation period could provide better embryo quality.

(Received March 15 2010)

(Accepted May 26 2010)

(Online publication January 14 2011)

Keywords:

• cycloheximide;
• in vitro fertilization;
• oocyte maturation;
• parthenogenetic activation;
• swine

Correspondence:

^c1 All correspondence to: Mariana Groke Marques. Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva no. 87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil. Tel: +55 11 30917916. Fax: +55 11 30917412. e-mail: marigroke@gmail.com