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Transfection of oocytes and other types of ovarian cells in rabbits after direct injection into uterine arteries of adenoviruses and plasmid/liposomes

Gene Therapy volume 10, pages 580–584 (2003)Cite this article

Abstract

Transfection of oocytes should be avoided in somatic gene therapy. However, several viral vectors including adenoviruses can transfect zona-pellucida-free eggs in vitro. During early stages of development, oocytes of postnatal ovaries lack the zona pellucida. Therefore, they may be susceptible to gene transfer and unintended toxic effects. The purpose of this study was to see whether the injection of adenoviruses (1 × 1010 PFU) or plasmid (500 μg)/DOTMA:DOPE (1:2) liposomes directly into uterine arteries in pregnant rabbits leads to transfection of oocytes and other types of ovarian cells. LacZ and herpes simplex virus thymidine kinase (HSV-TK) were used as transgenes. It was found that both adenovirus and plasmid vectors transfected oocytes at the primordial and primary follicle stage when they were not protected by the zona pellucida, whereas no transfection was seen in oocytes surrounded by the zona pellucida. Efficient transfection of corpus luteum and granulosa cells was also detected by adenoviral and plasmid vectors. Transfection of oocytes and other ovarian cells was verified by X-gal staining and laser microdissection, followed by PCR analysis. HSV-TK gene transfer, followed by ganciclovir treatment, led to destruction of a significant number of oocytes, whereas HSV-TK gene transfer alone did not lead to toxic effects. It is concluded that the presence of a high concentration of adenovirus or plasmid vectors via the uterine artery may lead to transfection of zona-pellucida-free oocytes and other ovarian cells.

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Acknowledgements

This study was supported by grants from Finnish Academy and Kuopio University Hospital (EVO grant 5130). We thank Ms Mervi Nieminen and Ms Anne Martikainen for technical assistance and Ms Marja Poikolainen for preparing the manuscript.

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Authors and Affiliations

  1. A.I. Virtanen Institute, Kuopio, Finland

    A Laurema, A Heikkilä, T Heikura, P Lehtolainen, T T Tuomisto & S Ylä-Herttuala

  2. Department of Gynecology and Obstetrics, University of Kuopio, Kuopio, Finland

    A Heikkilä, L Keski-Nisula & S Heinonen

  3. Department of Radiology, University of Kuopio, Kuopio, Finland

    H Manninen

  4. Department of Medicine, University of Kuopio, Kuopio, Finland

    S Ylä-Herttuala

  5. Gene Therapy Unit, Kuopio University Hospital, Kuopio, Finland

    S Ylä-Herttuala

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  1. A Laurema
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Laurema, A., Heikkilä, A., Keski-Nisula, L. et al. Transfection of oocytes and other types of ovarian cells in rabbits after direct injection into uterine arteries of adenoviruses and plasmid/liposomes. Gene Ther 10, 580–584 (2003). https://doi.org/10.1038/sj.gt.3301918

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  • DOI: https://doi.org/10.1038/sj.gt.3301918

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