African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12486

Full Length Research Paper

High efficacy of fibroblast and hepatocyte growth factors on the in vitro blastocyst production of post-thaw mouse two-cell embryo: The cryotop method

Fatemeh Ghasemian1, Mahnaz Azarnia2, Abtin Heidarzadeh3, Shervin Ghadarjani4 and Mohammad Hadi Bahadori5*
1Department of Biology, Faculty of Science, Kharazmi (Tarbiat moallem) University, Tehran, Iran. 2Department of Biology, Faculty of Science, Kharazmi (Tarbiat moallem) University, Tehran, Iran. 3Department of Community Medicine, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran. 4Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran. 5Cellular and Molecular Research Center, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran.  
Email: [email protected]

  •  Accepted: 05 September 2011
  •  Published: 30 November 2011

Abstract

There is great need to improve our understanding of what increases an embryo’s development potential, after vitrification-thawing processes. For this subject, 358 two-cell stage embryos were collected from oviduct of pregnant two-day old mice and vitrified. After thawing, embryos were cultured in Tyrode's (T6) medium supplemented with different doses of fibroblast growth factor (FGF; 0, 10, 20, 50 and 100 ng/ml) and hepatocyte growth factor (HGF; 0, 10, 20, 50 and 100 ng/ml) until the blastocyst stage. To determine quality of blastocysts, blastocysts were stained with hoechst and propidium iodide. After culture for 24 h, 92.65% of treated embryos with 20 ng/ml of FGF had higher (P<0.05) survival rate in comparison to the control group (84%). Blastocyst embryo formation rate were 79.43% (P< 0.01), and 67.46% (P< 0.05) in the treated groups with 20 ng/ml of FGF and 20 ng/ml of HGF, respectively, which were significantly different from the control (56%). In the treatment group with 20 ng/ml of FGF, blastocysts with >64 cells had a significantly higher inner cell mass (ICM) in comparison to the control group (P< 0.01). In conclusion, in this experiment, addition of growth factors in the culture had favorable effects on post-thawed cleavage of vitrified 2-cell embryos and blastocyst quality.

 

Key words: Blastocyst quality, growth factors, preimplantation development, vitrification.