title: A novel method for preserving cultured limbal epithelial cells.
abstract: OBJECTIVE
  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 degrees 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.
authors.affiliation: Department of Ophthalmology, Centre for Eye Research, University of Oslo, Ullevål University Hospital, Kirkeveien 166, 0407 Oslo, Norway.
authors.forename: Tor Paaske
authors.initials: TP
authors.lastname: Utheim
authors.forename: Sten
authors.initials: S
authors.lastname: Raeder
authors.forename: Øygunn Aass
authors.initials: ØA
authors.lastname: Utheim
authors.forename: Yiqing
authors.initials: Y
authors.lastname: Cai
authors.forename: Borghild
authors.initials: B
authors.lastname: Roald
authors.forename: Liv
authors.initials: L
authors.lastname: Drolsum
authors.forename: Torstein
authors.initials: T
authors.lastname: Lyberg
authors.forename: Bjørn
authors.initials: B
authors.lastname: Nicolaissen
mesh_terms: D000650:Amnion; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D003470:Culture Media; D006801:Humans; D016850:Limbus Corneae / Q000166:cytology*; D013696:Temperature
chemical_list: D003470:Culture Media