April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Comparison of Storage of Cultured Human Limbal Epithelial Cells in HEPES-MEM, Optisol-GS and PAA Quantum 286 for 4 days at 23°C
Author Affiliations & Notes
  • Tor P. Utheim
    Ophthalmology,
    Oslo University Hospital, Oslo, Norway
  • Øygunn A. Utheim
    Ophthalmology,
    Oslo University Hospital, Oslo, Norway
  • Jon R. Eidet
    Center for Clinical Research,
    Oslo University Hospital, Oslo, Norway
  • Maria de la Paz
    Institut Universitari Barraquer/Universitat Autonoma de Barcelona, Barcelona, Spain
  • Sten Raeder
    Ophthalmology, Stavanger University Hospital, Stavanger, Norway
  • Borghild B. Roald
    Pathology,
    Oslo University Hospital, Oslo, Norway
  • Torstein Lyberg
    Center for Clinical Research,
    Oslo University Hospital, Oslo, Norway
  • Footnotes
    Commercial Relationships  Tor P. Utheim, Patent applications on storage of epithelial cells have been filed (P); Øygunn A. Utheim, Patent applications on storage of epithelial cells have been filed (P); Jon R. Eidet, None; Maria de la Paz, None; Sten Raeder, Patent applications on storage of epithelial cells have been filed (P); Borghild B. Roald, None; Torstein Lyberg, Patent applications on storage of epithelial cells have been filed (P)
  • Footnotes
    Support  The Research Council of Norway, FORNY Grant
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5124. doi:
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      Tor P. Utheim, Øygunn A. Utheim, Jon R. Eidet, Maria de la Paz, Sten Raeder, Borghild B. Roald, Torstein Lyberg; Comparison of Storage of Cultured Human Limbal Epithelial Cells in HEPES-MEM, Optisol-GS and PAA Quantum 286 for 4 days at 23°C. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5124.

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

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Abstract

Purpose: : The feasibility of storage of cultured human limbal epithelial cells (HLEC) in serum-free media for as much as 4 days has not been demonstrated. Eye bank storage in a closed container enables important advantages: 1) transportation to eye clinics without culture facilities; 2) quality and microbiological assessment; and 3) increased flexibility in terms of operation time.

Methods: : 2-week HLEC cultures were stored in serum-free (HEPES-MEM and Optisol GS) or serum-containing (PAA Quantum 286) media for 4 days at 23°C in a closed container. Analyses included light microscopy, immunohistochemistry, and a calcein-acetoxymethyl ester/ethidium homodimer 1 viability assay. The Mann-Whitney test was applied for statistical evaluation.

Results: : Cultured HLEC stored in HEPES-MEM, Optisol-GS and PAA Quantum 286 for 4 days were well-preserved, attaching satisfactorily to the amniotic membranes with occasional intercellular edema. Basal layer viability of cultured HLEC prior to storage (98.1% ± 2.6%) was compared with viability after 4 days of storage in HEPES-MEM (97.3% ± 2.5%; p = 0.22), Optisol GS (91.5% ± 9.7%; p < 0.05), and PAA Quantum 286 (94.6% ± 2.5%; p < 0.01). The epithelial thickness prior to storage (29.0 µm ± 12.1 µm) was compared with the thickness after 4 days of storage in HEPES-MEM (21.2 µm ± 5.8 µm; p = 0.46), Optisol GS (15.4 µm ± 6.6 µm; p < 0.05), and PAA Quantum 286 (22.7 µm ± 16.0 µm; p = 0.32).

Conclusions: : The present study demonstrates successful storage of cultured HLEC in serum-free HEPES-MEM for 4 days. This may increase the availability of ocular surface stem cell therapy worldwide.

Keywords: cornea: epithelium • cornea: storage • anterior segment 
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