Cell and Tissue Research

, Volume 296, Issue 2, pp 371–383 | Cite as

Factors affecting proliferation and dedifferentiation of primary bovine oviduct epithelial cells in vitro

  • Judith Reischl
  • Katja Prelle
  • Heidrun Schöl
  • Christine Neumüller
  • Ralf Einspanier
  • Fred Sinowatz
  • E. Wolf
Regular Article

Abstract

 The oviduct is the physiological site for key events in reproduction, such as capacitation of spermatozoa, fertilization and early embryonic development. Interactions between oviduct epithelial cells and gametes or embryos cannot sufficiently be studied in vivo. Therefore, model systems are needed which mimic in vivo conditions most closely. In this study we optimised the method for isolating bovine oviduct cells and compared different cell support materials as well as two culture systems (perfusion vs static culture) for their ability to maintain characteristic morphological and functional features of oviduct cells. Out of nine different cell support materials tested, cellulose nitrate (0.45 µm pore size) was the most suitable to maintain cells in a manner similar to freshly isolated oviduct epithelial cells. Comparing static vs perfusion culture by electron microscopy, morphological differences of the cells were insignificant in the first days of culture, while they became more evident after 8 days. The cells in the static system lost typical characteristics such as columnar shape, cilia and secretory protrusions, while these features were still present in perfusion culture. In addition, intense ciliogenesis and cytoplasmic organelles for protein synthesis were found under perfusion conditions. These findings were underlined by differences in expression of the oviduct-specific oestrus-associated glycoprotein 85–97 kDa (GP 85–97) gene as revealed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The RNA levels of this specific gene were significantly higher in perfusion compared to the static culture system. Our data show clear advantages of perfusion vs static culture for primary bovine oviduct epithelial cells.

Key words Oviduct epithelium Perfusion culture Cell support Permeable membranes Reverse transcription-polymerase chain reaction Bovine 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Judith Reischl
    • 1
  • Katja Prelle
    • 1
  • Heidrun Schöl
    • 3
  • Christine Neumüller
    • 4
  • Ralf Einspanier
    • 2
  • Fred Sinowatz
    • 4
  • E. Wolf
    • 1
  1. 1.Department of Molecular Animal Breeding and Genetics, Ludwig-Maximilian University, Hackerstr. 27, D-85764 Oberschleissheim, Munich, Germany e-mail: ewolf@lmb.uni-muenchen.de; Tel.: +49 89 74017 274; Fax: +49 89 74017 368DE
  2. 2.Institute of Physiology, Technical University Munich-Weihenstephan, Freising, GermanyDE
  3. 3.Institute for Comparative Tropical Medicine and Parasitology, Ludwig-Maximilian University, Munich, GermanyDE
  4. 4.Institute of Veterinary Anatomy II, Ludwig-Maximilian University, Munich, GermanyDE

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