The Effects of Epithelial Viability on Stromal Keratocyte Apoptosis in Porcine Corneas Stored in Optisol-GS

Chang, Shu‐Wen; Wang, Yao‐Horng; Pang, Jong‐Hwei S.

doi:10.1097/01.ico.0000160970.58330.09

Cited by 10 publications

(8 citation statements)

References 33 publications

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“…Optisol GS™ is a common preservative medium used for corneal tissues, and has been shown to be particularly suitable for preservation of the corneal endothelium in hypothermia36; however, some reports have suggested that the medium is not suitable for the preservation of corneal epithelium3738. The results of our experiment clearly showed that Optisol GS™ is not suitable for preservation of the corneal epithelium or its stem cells.…”

Section: Discussioncontrasting

confidence: 53%

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Katori

Hayashi

Kobayashi

et al. 2016

Sci Rep

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Clinical trials have been performed using autologous tissue-engineered epithelial cell sheets for corneal regenerative medicine. To improve stem cell-based therapy for convenient clinical practice, new techniques are required for preserving reconstructed tissues and their stem/progenitor cells until they are ready for use. In the present study, we screened potential preservative agents and developed a novel medium for preserving the cell sheets and their stem/progenitor cells; the effects were evaluated with a luciferase-based viability assay. Nrf2 activators, specifically ebselen, could maintain high ATP levels during preservation. Ebselen also showed a strong influence on maintenance of the viability, morphology, and stem cell function of the cell sheets preserved under hypothermia by protecting them from reactive oxygen species-induced damage. Furthermore, ebselen drastically improved the preservation performance of human cornea tissues and their stem cells. Therefore, ebselen shows good potential as a useful preservation agent in regenerative medicine as well as in cornea transplantation.

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Section: Discussioncontrasting

confidence: 53%

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Katori

Hayashi

Kobayashi

et al. 2016

Sci Rep

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“…However, some reports have suggested that the use of Optisol-GS is not suitable for the preservation of corneal epithelium 8,9 . In contrast, there are other solutions that have been shown to be an effective compound for the preservation of human corneal epithelium 10 .…”

Section: Discussionmentioning

confidence: 99%

Comparison of rabbit corneal changes during different preservation techniques using Optisol-GS and airlift.

Tang

Uematsu

Harada

et al. 2023

Preprint

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Background: A previous study has suggested that airlift condition is superior to Optisol-GS condition in preserving the limbal tissue of human cornea. Purpose: To investigate a new preservation device that preserves the cornea while separating epithelial and endothelial areas. We compared the differences after preserving the corneal epithelium in different conditions. Methods: 24 corneas of New Zealand rabbits were divided into 4 groups in which the corneal epithelia submersed in Optisol-GS or under airlift conditions for 1 and 2 weeks, (4°C). Transparency, OCT, H&E staining and epithelial migration tests were used to assess the corneal status. Results: The epithelial migration examination showed statistical difference indicating that the epithelium had larger migration ability after airlift conditions. Corneas in the 1-week Optisol-GS group were the most transparent, followed by the 1-week airlift group. OCT showed progressive increase of corneal thickness till the end of the study. H&E staining results showed that the epithelial cells retained intact cellular structure and morphology of the cells for both the 1-week preserved groups. However, there was disruption of the corneal epithelial cell structure for both 2-week preserved groups. Conclusion: Corneal epithelium preserved under hypothermic airlift conditions was comparable to that found for Optisol-GS conditions.

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“…Furthermore, CSK preservation in Optisol-GS was reported to be suboptimal. The cells underwent apoptosis, which was more significant after 5 days of storage in Optisol-GS (2). Debridement of corneal epithelium during storage further boosted CSK apoptosis.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were seeded at 10 4 cells per cm 2 on collagen I-coated culture wells (BD Biosciences, Franklin Lakes, NJ, USA). To propagate genuine CSKs, our results must meet three basic requirements: (1) maintenance of cells with dendritic morphology, (2) lack of fibroblast features (negative expression of αSMA and F-actin stress fiber pattern), and (3) expression of CSK markers (ALDH1A1, ALDH3A1, lumican, keratocan, and COL8A2). We cultured the cells with various chemicals and growth factors, including soluble amnion stromal extract (ASE; preparation details as shown below), ROCK inhibitor Y27632 (10 mM; Millipore, Billerica, MA, USA), insulin-like growth factor 1 (IGF1; 10 ng/ml; Invitrogen), and/or fetal bovine serum (FBS; Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

Ex Vivo Propagation of Human Corneal Stromal “Activated Keratocytes” for Tissue Engineering

et al. 2015

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Keratoconus is a corneal disorder characterized by a thinning of stromal tissue, and the affected patients have induced astigmatism and visual impairment. It is associated with a loss of corneal stromal keratocytes (CSKs). Hence, reconstructing stromal tissue with autologous CSK replacement can be a viable alternative to corneal transplantation, which is restricted by the global donor material shortage and graft rejection. Human CSKs are normally quiescent and express unique markers, like aldehyde dehydrogenases and keratocan. In serum culture, they proliferate, but lose their characteristic phenotype and become stromal fibroblasts. Here we report a novel culture cocktail to ex vivo propagate and maintain CSKs. Primary human CSKs were obtained from adult donors and cultured with soluble human amnion stromal extract (ASE), rho-associated coiled-coil-forming protein serine/threonine kinase inhibitor Y-27632, and insulin-like growth factor-1 (collectively named as ERI). Protein profiling using mass spectrometry followed by MetaCore™ pathway analysis predicted that ASE proteins might participate in transforming growth factor-β (TGF-β) signaling and fibroblast development, cell adhesion, extracellular matrix remodeling, and immune response. In culture with 0.5% fetal bovine serum and ERI, the population of "activated keratocytes" was expanded. They had much lowered expression of both keratocyte and fibroblast markers, suppressed TGF-β-mediated Smad2/3 activation, and lacked fibroblast-mediated collagen contractibility. These "activated keratoctyes" could be propagated for six to eight passages ex vivo, and they regained CSK-specific dendritic morphology and gene marker expression, including aldehyde dehydrogenases, lumican, and keratocan biosynthesis, expression, and secretion when returned to serum-depleted ERI condition. This novel cocktail maintained human CSKs in both adherent and suspension cultures with proper keratocyte features and without the transformation to stromal fibroblasts. Thus, human CSKs can be ex vivo propagated as transient "activated keratocytes." This could provide sufficient number of genuine CSKs for corneal tissue engineering.

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The Effects of Epithelial Viability on Stromal Keratocyte Apoptosis in Porcine Corneas Stored in Optisol-GS

Abstract: Stromal keratocytes underwent apoptosis in Optisol-GS. The absence of corneal epithelium during preservation further increased the stromal keratocyte apoptosis.

Cited by 10 publications

References 33 publications

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Ebselen Preserves Tissue-Engineered Cell Sheets and their Stem Cells in Hypothermic Conditions

Comparison of rabbit corneal changes during different preservation techniques using Optisol-GS and airlift.

Ex Vivo Propagation of Human Corneal Stromal “Activated Keratocytes” for Tissue Engineering

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