Immunocytochemical Diagnosis of Limbal Stem Cell Deficiency

Poli, Muriel; Burillon, C.; Auxenfans, Céline; Rovere, M.R.; Damour, O.

doi:10.1097/ico.0000000000000457

Cited by 25 publications

(17 citation statements)

References 27 publications

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“…It seems a good approach to distinguish the limbus from the conjunctiva based on goblet cells, which were used to diagnose LSCD [ 109 ]. Controversially, the Stevens–Johnsons syndrome and long-term administration of glaucoma drugs could lead to LSCD with no goblet cells invasion [ 110 ]. Some investigators reported that CK19, CK13 and MUC5AC can serve as biomarkers of conjunctival epithelial cells [ 110 , 111 ], but others argued that the expression of CK19 was detected both on the limbal and CECs [ 27 ].…”

Section: Difficulties In the Study Of Lsc-specific Biomarkersmentioning

confidence: 99%

“…Controversially, the Stevens–Johnsons syndrome and long-term administration of glaucoma drugs could lead to LSCD with no goblet cells invasion [ 110 ]. Some investigators reported that CK19, CK13 and MUC5AC can serve as biomarkers of conjunctival epithelial cells [ 110 , 111 ], but others argued that the expression of CK19 was detected both on the limbal and CECs [ 27 ]. Although Ramirez-Miranda et al believed that CK13 was more specific than CK19 in terms of identification of conjunctival epithelial cells [ 92 ], Poli et al showed that CK13 was expressed in both the conjunctival epithelium and the suprabasal and superficial layers of the limbal epithelium [ 110 ].…”

Section: Difficulties In the Study Of Lsc-specific Biomarkersmentioning

confidence: 99%

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An Insight into the Difficulties in the Discovery of Specific Biomarkers of Limbal Stem Cells

Guo

Zhang

Jia

et al. 2018

IJMS

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Keeping the integrity and transparency of the cornea is the most important issue to ensure normal vision. There are more than 10 million patients going blind due to the cornea diseases worldwide. One of the effective ways to cure corneal diseases is corneal transplantation. Currently, donations are the main source of corneas for transplantation, but immune rejection and a shortage of donor corneas are still serious problems. Graft rejection could cause transplanted cornea opacity to fail. Therefore, bioengineer-based corneas become a new source for corneal transplantation. Limbal stem cells (LSCs) are located at the basal layer in the epithelial palisades of Vogt, which serve a homeostatic function for the cornea epithelium and repair the damaged cornea. LSC-based transplantation is one of the hot topics currently. Clinical data showed that the ratio of LSCs to total candidate cells for a transplantation has a significant impact on the effectiveness of the transplantation. It indicates that it is very important to accurately identify the LSCs. To date, several putative biomarkers of LSCs have been widely reported, whereas their specificity is controversial. As reported, the identification of LSCs is based on the characteristics of stem cells, such as a nuclear-to-cytoplasm ratio (N/C) ≥ 0.7, label-retaining, and side population (SP) phenotype. Here, we review recently published data to provide an insight into the circumstances in the study of LSC biomarkers. The particularities of limbus anatomy and histochemistry, the limits of the current technology level for LSC isolation, the heterogeneity of LSCs and the influence of enzyme digestion are discussed. Practical approaches are proposed in order to overcome the difficulties in basic and applied research for LSC-specific biomarkers.

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Section: Difficulties In the Study Of Lsc-specific Biomarkersmentioning

confidence: 99%

Section: Difficulties In the Study Of Lsc-specific Biomarkersmentioning

confidence: 99%

An Insight into the Difficulties in the Discovery of Specific Biomarkers of Limbal Stem Cells

Guo

Zhang

Jia

et al. 2018

IJMS

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“…Severe damage to the limbal epithelial cells from various etiologies in the limbal region may lead to loss of the limbal epithelial cells [ 2 ], so called limbal stem cell deficiency (LSCD). LSCD, manifested by chronic inflammation, neovascularization, and goblet cell invasion into the cornea, may be complicated by persistent corneal epithelial defects, ulceration, and even perforation of the cornea [ 3 , 4 ]. The cornea may ultimately be healed by fibrosis, however, the vision will be greatly impaired.…”

Section: Introductionmentioning

confidence: 99%

Y-27632, a ROCK Inhibitor, Promoted Limbal Epithelial Cell Proliferation and Corneal Wound Healing

et al. 2015

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Transplantation of ex vivo cultured limbal epithelial cells is proven effective in restoring limbal stem cell deficiency. The present study aimed to investigate the promoting effect of Y-27632 on limbal epithelial cell proliferation. Limbal explants isolated from human donor eyes were expanded three weeks on culture dishes and outgrowth of epithelial cells was subsequently subcultured for in vitro experiments. In the presence of Y-27632, the ex vivo limbal outgrowth was accelerated, particularly the cells with epithelial cell-like morphology. Y-27632 dose-dependently promoted the proliferation of in vitro cultured human limbal epithelial cells as examined by phase contrast microscopy and luminescent cell-viability assay 30 hours after the treatment. The colony forming efficacy determined 7 days after the treatment was enhanced by Y-27632 also in a dose-dependent manner. The number of p63- or Ki67-positive cells was dose-dependently increased in Y-27632-treated cultures as detected by immunofluorescent staining and western blotanalysis. Cell cycle analysis by flow cytometric method revealed an increase in S-phase proliferating cells. The epithelial woundclosure rate was shown to be faster in experimental group received topical treatment withY-27632 than the sham control using a rat corneal wounding model. These resultsdemonstrate that Y-27632 can promote both the ex vivo and in vitro proliferation oflimbal epithelial cell proliferation. The in vivo enhanced epithelial wound healingfurther implies that the Y-27632 may act as a new strategy for treating limbal stem cell deficiency.

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“…159 The directed differentiation of iPSCs into CECs depends on the expression of cytokeratins (CK) 12 and 13, while CK3 expression was evident in the cell lines derived from the CE. 160,161 To improve the yields of mature CECs and to obtain a stratified cell sheet resembling the native CE, a consistent and efficient stratification method has to be employed. It is not uncommon to detect variation in the differentiation potential among different hiPSC lines, with donor identity and gender being among the potential sources of variation in the case of hiPSC lines.…”

Section: Induced Pluripotent Stem Cells (Ips)mentioning

confidence: 99%