A novel method for preserving cultured limbal epithelial cells
- Tor Paaske Utheim1,
- Sten Raeder1,
- Øygunn Aass Utheim1,
- Yiqing Cai2,
- Borghild Roald3,
- Liv Drolsum1,
- Torstein Lyberg4,
- Bjørn Nicolaissen1
- 1Department of Ophthalmology, Centre for Eye Research, University of Oslo, Ullevål University Hospital, Oslo, Norway
- 2Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
- 3Department of Pathology, Ullevål University Hospital, University of Oslo, Oslo, Norway
- 4Centre for Clinical Research, Ullevål University Hospital, Oslo, Norway
- Correspondence to: MrS Raeder Department of Ophthalmology, Centre for Eye Research, University of Oslo, Ullevål University Hospital, Kirkeveien 166, 0407 Oslo, Norway;
- Accepted 2 November 2006
- Published Online First 23 November 2006
Aim: To investigate organ culture preservation of cultured limbal epithelial cells in order to enhance the availability of tissue-engineered epithelia that are used to treat patients with limbal stem cell deficiency.
Methods: Limbal epithelial cells were cultured for 3 weeks on intact amniotic membrane fastened to a polyester membrane carrier. The cultured epithelia were stored for 1 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 with non-preserved epithelia. In general, the cell borders were maintained, the nuclei showed 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 1 week at room temperature, while maintaining the original layered structure and undifferentiated phenotype.
Funding: This work was supported in part by the Eastern Norway Regional Health Authority, the Norwegian Association of the Blind and Partially Sighted, and the Blindmission IL.
Competing interests: None declared.