Transfection of EGFP to bovine spermatogonial colony through lipofection

Authors

1 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran-Iran

2 Department of Biotechnology, Iranian Research Organization for Sciences and Technology, Tehran-Iran

Abstract

 
BACKGROUND: Spermatogonial Stem Cells (SSCs) are the only stem cells in adults that can transfer genetic information to future generations. Considering that a single SSC gives rise to a vast number of spermatozoa, genetic manipulation of these cells is a potential novel technology with practical application to various animal species. OBJECTIVES: The aim of this study was to evaluate Enhanced Green Fluorescent Protein (EGFP) gene transfection into bovine spermatogonial colonies via liposome carrier and assess the best incubation day in uptake exogenous gene by spermatogonial colonies. METHODS: Transfection efficiency EGFP gene through lipofection was determined different in three days (day 4, 6 and 8) after the beginning of the culture by fluorescent microscope. Immunofluorescent staining against OCT4 and vimentin led to the confirmation of the nature of both SSCs and sertoli cells. RESULTS: Results showed that the transfected colonies through lipofection increased significantly (p<0.05) in each three days of transfection in comparison with those of the control groups. The transfection colonies were higher (significant) in comparision with those of the free exogenous gene carrier groups. The rate of infected colonies was higher when transfection proceed day 4. CONCLUSIONS: It was concluded that lipofectamine can be used safely for direct loading of exogenous DNA to spermatogonila colony, particularly during the fourth day of culture.
 

Keywords

References

 
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