May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Storage of Cultured Limbal Epithelial Cells at 23ºC and 31ºC in Organ Culture Medium and at 5ºC in Optisol-GS: A Light Microscopy and Phenotypic Study
Author Affiliations & Notes
  • T. Utheim
    Ullevål University Hospital, Oslo, Norway
    Department of Ophthalmology,
  • S. Raeder
    Ullevål University Hospital, Oslo, Norway
    Department of Ophthalmology,
  • Ø. Utheim
    Ullevål University Hospital, Oslo, Norway
    Department of Ophthalmology,
  • Y. Cai
    Department of Oral Biology, Faculty of Dentistry, University of Oslo, Norway
  • B. Roald
    Ullevål University Hospital, Oslo, Norway
    Department of Pathology,
  • T. Noren
    Ullevål University Hospital, Oslo, Norway
    Department of Pathology,
  • K. Haug
    Ullevål University Hospital, Oslo, Norway
    Department of Ophthalmology,
  • L. Drolsum
    Ullevål University Hospital, Oslo, Norway
    Department of Ophthalmology,
  • T. Lyberg
    Ullevål University Hospital, Oslo, Norway
    Center for Clinical Research,
  • B. Nicolaissen
    Ullevål University Hospital, Oslo, Norway
    Department of Ophthalmology,
  • Footnotes
    Commercial Relationships T. Utheim, None; S. Raeder, None; Ø. Utheim, None; Y. Cai, None; B. Roald, None; T. Noren, None; K. Haug, None; L. Drolsum, None; T. Lyberg, None; B. Nicolaissen, None.
  • Footnotes
    Support 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.
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 460. doi:
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      T. Utheim, S. Raeder, Ø. Utheim, Y. Cai, B. Roald, T. Noren, K. Haug, L. Drolsum, T. Lyberg, B. Nicolaissen; Storage of Cultured Limbal Epithelial Cells at 23ºC and 31ºC in Organ Culture Medium and at 5ºC in Optisol-GS: A Light Microscopy and Phenotypic Study. Invest. Ophthalmol. Vis. Sci. 2007;48(13):460.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose:: To compare 31°C organ culture storage and hypothermic storage of cultured limbal epithelial cells with our novel and recently reported method using 23°C organ culture preservation.

Methods:: Limbal epithelial cells were cultured for three weeks on amniotic membranes fastened to polyester membrane carriers. The cultured epithelia were stored for one week at 31°C in organ culture medium and at 5ºC in Optisol-GS and subsequently examined using light microscopy and immunohistochemistry.

Results:: At 23 ºC the cell borders were maintained, and the epithelia attached well to the amniotic membrane. Mild intercellular oedema was occasionally observed. In contrast, at both 31ºC and 5 ºC storage, detachment of epithelial cells and enlarged intercellular spaces occurred regularly. Pronounced detachment of the basal layer was observed at 5 ºC storage. Occurrence of fibroblasts was disclosed in 3/8 of the epithelia stored at 31ºC. Expression of p63, K19, K5, and vimentin was maintained in all groups. K3 was not expressed in either of the experimental groups. Furthermore, Integrin beta-1, E-cadherin, Connexin 43, K14, and Ki 67 demonstrated minor changes in immunostaining in the three groups.

Conclusions:: 31ºC organ culture storage and in particular hypothermic storage were less satisfactory in preserving cultured limbal epithelial cells than organ culture preservation at 23ºC. However, the original undifferentiated phenotype was maintained irrespective of the organ culture medium and temperature.

Keywords: cornea: basic science • cornea: epithelium • cornea: storage 
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