rss
Br J Ophthalmol 2009;93:980-983 doi:10.1136/bjo.2008.149591
  • Laboratory science
    • Original Article

Sterility control and long-term eye-bank storage of cultured human limbal epithelial cells for transplantation

  1. T P Utheim1,2,
  2. S Raeder1,2,
  3. Ø A Utheim2,
  4. M de la Paz3,
  5. B Roald4,
  6. T Lyberg1
  1. 1
    Center for Clinical Research, Ulleval University Hospital, Oslo, Norway
  2. 2
    Department of Ophthalmology, University of Oslo, Ulleval University Hospital, Norway
  3. 3
    Institut Universitari Barraquer/Universitat Autonoma de Barcelona, Barcelona, Spain
  4. 4
    Department of Pathology, Ulleval University Hospital, University of Oslo, Norway
  1. Dr S Raeder, Center for Clinical Research, University of Oslo, Ulleval University Hospital, Kirkeveien 166, 0407 Oslo, Norway; sten.rader{at}medisin.uio.no
  • Accepted 13 December 2008
  • Published Online First 11 February 2009

Abstract

Background/aims: To assess sterility of cultured human limbal epithelial cells (HLEC) and to investigate the viability, morphology and phenotype of cultured HLEC following 2 and 3 weeks of organ culture storage.

Methods: HLEC cultured on amniotic membranes were stored in organ culture medium in a closed container at 23°C. Sterility of storage media was tested using a Bactec 9240 blood culture instrument (Becton Dickinson, Maryland) for incubation and periodic reading. Viability was analysed by calcein-acetoxymethyl ester/ethidium homodimer-1 assay, morphology by light microscopy and cellular phenotype by immunohistochemistry.

Results: No microbial contamination was observed after 1 week’s storage. Viability of cultured HLEC was 87.9 (SD 6.4)% and 52.7 (13.1)% after 2 and 3 weeks of storage, respectively, compared with 98.8 (2.6)% before storage (p<0.001). The multilayered structure was preserved in 70% of cultures following 2 weeks of storage but lost after 3 weeks. A less differentiated phenotype was maintained.

Conclusion: This study is the first to verify the sterility of HLEC cultures prior to transplantation. Although a slight decrease in viability was observed following 2 weeks of storage, the HLEC sheets remain acceptable, whereas 3 week’s storage was unsatisfactory.

Footnotes

  • Competing interests: TPU and SR have filed a patent application on storage of cultured limbal epithelial cells.

  • Funding: This work was supported by the South-Eastern Norway Regional Health Authority.

  • Ethics approval: Ethics approval was provided by Regional komite for etikk, Region Øst-Norge.

  • Patient consent: Obtained.

This Article

  1. Abstract
  2. Full text
  3. PDF
  4. All versions of this article:
    1. bjo.2008.149591v1
    2. 93/7/980 most recent

Responses

  1. Submit a response
  2. No responses published

Register for free content

The full back archive is now available for all BMJ Journals. Institutional subscribers may access the entire archive as part of their subscription. Personal subscribers will also have access to all content when logged in. Non-subscribers who register have free access to all articles published before 2006 right back to volume 1 issue 1. Register here to access the free archive of all BMJ Journals.

Don't forget to sign up for content alerts so you keep up to date with all the articles as they are published.