ArticlePloS one2017
Optimizing the role of limbal explant size and source in determining the outcomes of limbal transplantation: An in vitro study.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 35 citations in OpenAlex.
- Review
- Substrate engineering using naturally biomimicking corneal cell topography for preserving stemness of corneal limbal epithelial-stem cells.Iranian journal of basic medical sciences · 2025Article
- Targeting limbal epithelial stem cells: master conductors of corneal epithelial regeneration from the bench to multilevel theranostics.Journal of translational medicine · 2024Review
- Focus on limbal stem cell deficiency and limbal cell transplantation.World journal of transplantation · 2023Review
- Preparation of human amniotic membrane for transplantation in different application areas.Frontiers in transplantation · 2023Review
- Limbal epithelial stem cell sheets from young donors have better regenerative potential.Scientific reports · 2022Article
- Clinical, histological and immunohistochemistry characteristics of cornea in the sequelae stage of chronic vernal keratoconjunctivitis.Indian journal of ophthalmology · 2022Article
- Review
- Current perspectives of limbal-derived stem cells and its application in ocular surface regeneration and limbal stem cell transplantation.Stem cells translational medicine · 2021Review
- Clinical outcomes of modified simple limbal epithelial transplantation for limbal stem cell deficiency in Chinese population: a retrospective case series.Stem cell research & therapy · 2021Article
- Simple limbal epithelial transplantation: Current status and future perspectives.Stem cells translational medicine · 2020Review
- Amniotic Membrane Transplantation in Ophthalmology: An Updated Perspective.Clinical ophthalmology (Auckland, N.Z.) · 2020Review
- Characterization of limbal explant sites: Optimization of stem cell outgrowth in in vitro culture.PloS one · 2020Article
- Simple limbal epithelial transplantation (SLET): Review of indications, surgical technique, mechanism, outcomes, limitations, and impact.Indian journal of ophthalmology · 2019Review
- The application of human amniotic membrane in the surgical management of limbal stem cell deficiency.The ocular surface · 2019Review
- Article
- Twenty years of limbal epithelial therapy: an update on managing limbal stem cell deficiency.BMJ open ophthalmology · 2018Review
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeSimple limbal epithelial transplantation (SLET) and cultivated limbal epithelial transplantation (CLET) are proven clinical techniques for treating limbal stem cell deficiency (LSCD). However, the ideal size and number of the limbal explants required for transplantation has not been clearly elucidated. This in vitro study aimed to determine the optimal limbal explant size required for complete corneal epithelialization by characterizing the cell expansion.
methodsLimbal explants obtained from both live and cadaveric biopsies were cultured on the denuded amniotic membrane. Explant size and the explant cell outgrowth (expansion) were measured using ImageJ software with respect to days. Cultures were characterized by assessing the rate of proliferation of cells with 5-bromo-2'-deoxyuridine (BrdU) assay along with the expression of different stem cell markers (ABCG2, p63α), corneal epithelial (CK3+12) and adherens junction molecules (E-Cadherin) by immunofluorescence.
resultsExplants from live biopsies had 80% growth potential in vitro whereas 40% of the cadaveric tissue failed to grow. Minimum explant sizes of 0.3 mm2 for live and ≥0.5 mm2 for cadaveric tissue had a mean expansion areas of 182.39±17.06 mm2 and 217.59±16.91 mm2 respectively suggesting adequate growth potential of the explants. Mean total percentage of proliferative cells was 31.80±3.81 in live and 33.49±4.25 in cadaveric tissue expansion. The expression was noted to be similar in cells cultured from cadaveric compared to cells cultured from live limbal tissue with respect to ABCG2, p63α, CK(3+12) and E-cadherin.
conclusionOur findings show that a minimal amount of 0.3 mm2 live tissue would be sufficient for ample limbal cell expansion in vitro. Cadaveric explants <0.5 mm2 had poor growth potential. However, larger explants (≥ 0.5 mm2) had growth rate and proliferative potential similar to the live tissue. These findings could prove to be critical for clinical success especially while attempting cadaveric limbal transplantation. This study provides a novel clinical strategy for enhancing efficacy of the limbal transplantation surgery and opens the probability of even using the cadaveric tissue by considering the size of explant.
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