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A novel method for preserving cultured limbal epithelial cells
  1. Tor Paaske Utheim,
  2. Sten Raeder (sten.rader{at}medisin.uio.no),
  3. Øygunn Aass Utheim,
  4. Yiqing Cai,
  5. Borghild Roald,
  6. Liv Drolsum,
  7. Torstein Lyberg,
  8. Bjørn Nicolaissen
  1. Center for Eye Research, University of Oslo, Department of Ophthalmology, Norway
  2. Center for Eye Research, University of Oslo, Department of Ophthalmology, Norway
  3. Center for Eye Research, University of Oslo, Department of Ophthalmology, Norway
  4. Department of Oral Biology, Faculty of Dentistry, University of Oslo, Norway
  5. Department of Pathology, Ullevål University Hospital, University of Oslo, Norway
  6. Center for Eye Research, University of Oslo, Department of Ophthalmology, Norway
  7. Center for Clinical Research, Ullevål University Hospital, Norway
  8. Center for Eye Research, University of Oslo, Department of Ophthalmology, Norway

    Abstract

    Background/aims: To investigate organ culture preservation of cultured limbal epithelial cells in order to enhance the availability of tissue engineered epithelia used to treat patients with limbal stem cell deficiency.

    Methods: Limbal epithelial cells were cultured for three weeks on intact amniotic membrane fastened to a polyester membrane carrier. The cultured epithelia were stored for one week at 23°C in organ culture medium. The preserved epithelia were then examined using a colorimetric cell viability assay, light microscopy and immunohistochemistry.

    Results: The viability of the preserved epithelia was 84±20%, and no statistically significant difference was found compared to non-preserved epithelia. In general, the cell borders were maintained, the nuclei demonstrated no sign of degeneration, and the original layered structure was preserved. Mild intercellular oedema was occasionally observed. Expression of p63, K19 and vimentin was maintained.

    Conclusions: Cultured limbal epithelial cells can be preserved in organ culture medium for one week at room temperature while maintaining the original layered structure and undifferentiated phenotype.

    • corneal epithelium
    • limbal stem cells
    • organ culture preservation
    • tissue engineering

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