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Bovine somatic cell nuclear transfer using mitomycin C-mediated chemical oocyte enucleation

Published online by Cambridge University Press:  30 April 2019

M.T. Moura*
Affiliation:
Embrapa Recursos Genéticos e Biotecnologia, 70770–917, Brasília-DF, Brazil Universidade de Brasília, 70910–900, Brasília-DF, Brazil
R.V. Sousa
Affiliation:
Embrapa Recursos Genéticos e Biotecnologia, 70770–917, Brasília-DF, Brazil
C.M. Lucci
Affiliation:
Universidade de Brasília, 70910–900, Brasília-DF, Brazil
R. Rumpf
Affiliation:
Embrapa Recursos Genéticos e Biotecnologia, 70770–917, Brasília-DF, Brazil Universidade de Brasília, 70910–900, Brasília-DF, Brazil
*
Address for correspondence: Marcelo Tigre Moura. Federal University of São Paulo-UNIFESP, Diadema-SP, Brazil. Tel: +55 11 4044 0500 extension 3474. E-mail: [email protected]

Summary

Chemical oocyte enucleation holds the potential to ease somatic cell nuclear transfer (SCNT), although high enucleation rates remain limited to micromanipulation-based approaches. Therefore, this study aimed to test mitomycin C (MMC) for use in bovine functional chemical oocyte enucleation. Incubation of denuded eggs in 10 µg ml−1 MMC for different periods did not affect most maturation rates (0.5 h: 85.78%A, 1.0 h: 72.77%B, 1.5 h: 83.87%A, and 2.0 h: 82.05%A) in comparison with non-treated controls (CTL; 85.77%A). Parthenogenetic development arrest by MMC was efficient at cleavage (CTL: 72.93%A, 0.5 h: 64.92%A,B, 1.0 h: 60.39%B,C, 1.5 h: 66.35%A,B, and 2.0 h: 53.84%C) and blastocyst stages (CTL: 33.94%A, 0.5 h: 7.58%B, 1.0 h: 2.47%C, 1.5 h: 0.46%C, and 2.0 h: 0.51%C). Blastocysts were obtained after nuclear transfer (NT) using MMC enucleation [NT(MMC): 4.54%B] but at lower rates than for the SCNT control [NT(CTL): 26.31%A]. The removal of the meiotic spindle after MMC incubation fully restored SCNT blastocyst development [NT(MMC+SR): 24.74%A]. Early pregnancies were obtained by the transfer of NT(MMC) and NT(MMC+SR) blastocysts to synchronized recipients. In conclusion, MMC leads to functional chemical oocyte enucleation during SCNT and further suggests its potential for application towards technical improvements.

Type
Research Article
Copyright
© Cambridge University Press 2019 

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Footnotes

Current address: Federal University of São Paulo-UNIFESP, 09972–270, Diadema-SP, Brazil.

Current address: Geneal Genética e Biotecnologia Animal, 38020-970, Uberaba-MG, Brazil.

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