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    Establishment and characterization of an adherent pure epithelial cell line derived from the bovine oviduct (2008)

    Art
    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Autoren
    Schoen, J
    Bondzio, A
    Topp, K
    Einspanier, R
    Quelle
    Theriogenology : an international journal of animal reproduction
    Bandzählung: 69
    Heftzählung: 5
    Seiten: 536 – 545
    ISSN: 0093-691x
    Sprache
    Englisch
    Verweise
    DOI: 10.1016/j.theriogenology.2007.10.024
    Pubmed: 18242684
    Kontakt
    Institut für Veterinär-Biochemie

    Oertzenweg 19 b
    14163 Berlin
    +49 30 838 62225
    biochemie@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    The oviduct in vivo has to perform various tasks: maturation and transport of the gametes, milieu preparation for fertilization and embryonic development, and transport of the embryo. The complex arrangement of endocrine and paracrine signals being exchanged between the early embryo and the inner cell layers of the oviduct is barely understood. Therefore, a reproducible, well-characterized oviduct epithelial cell line as well as an optimized transfection protocol for DNA vectors and siRNA for this cell line has been established. A bovine oviduct primary cell culture system has been optimized using a selection medium permitting the survival of only epithelial cells. From this we established an adherent bovine oviduct pure epithelial cell line (aBOPEC-1). This cell line maintains some important characteristics of the primary cells such as the expression of estrogen receptors and p450 aromatase but it lacks some characteristics due to the selection and dedifferentiation processes (cilia, expression of progesterone receptor and oviduct specific glycoprotein-1). Optimization of the transfection protocols finally revealed a suitable DNA-transfection procedure yielding transfection efficiencies of over 50%. Additionally, siRNA transfection efficiency reached more than 90%. This new cell line builds an essential basis especially for future functional studies in the oviduct epithelium using distinct knock down experiments.