Challenges in Corneal Endothelial Cell Culture

Wongvisavavit, Rintra; Parekh, Mohit; Ahmad, Sajjad; Daniels, Julie T.

doi:10.2217/rme-2020-0202

Cited by 21 publications

(11 citation statements)

References 113 publications

(180 reference statements)

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“…The late-onset cells appear to originate from the far peripheral area of the endothelium, a region that has been referred to as a progenitor enriched region (TZ) with the potential to generate mature human corneal EC [ 27 , 28 ]. For the far peripheral endothelium a high mitogenic activity has been reported due to their propensity to sphere formation when cultured on low adhesion surfaces [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

Early and late-onset cell migration from peripheral corneal endothelium

et al. 2023

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In this study we describe peripheral corneal endothelial cell migration in vitro in the absence and presence of a ROCK-inhibitor. For this study, 21 corneal endothelial graft rims, with attached trabecular meshwork (TM), were prepared from Descemet membrane-endothelial cell sheets by 6.5 mm trepanation. For the initial proof-of-concept, 7 outer graft rims were cultured in a thermo-reversible hydrogel matrix for up to 47 days. To assess the effect of a ROCK-inhibitor, 14 paired outer rims were cultured either with or without ROCK-inhibitor for up to 46 days. At the end of culture, tissue was retrieved from the hydrogel matrix and examined for cell viability and expression of different endothelial cell markers (ZO-1, Na+/K+-ATPase, NCAM, glypican, and vimentin). All cultured rims remained viable and displayed either single regions (n = 5/21) or collective areas (n = 16/21) of cell migration, regardless of the presence or absence of ROCK-inhibition. Migration started after 4±2 days and continued for at least 29 days. The presence of ROCK-inhibitor seemed to contribute to a more regular cell morphology of migrating cells. In addition, 7 outer rims demonstrated a phenotypically distinct late-onset but fast-growing cell population emerging from the area close to the limbus. These cells emerged after 3 weeks of culture and appeared less differentiated compared to other areas of migration. Immunostaining showed that migrated cells maintained the expression patterns of endothelial cell markers. In conclusion, we observed 2 morphologically distinct migrating cell populations with the first type being triggered by a broken physical barrier, which disrupted contact inhibition and the second, late-onset type showing a higher proliferative capacity though appearing less differentiated. This cell subpopulation appeared to be mediated by stimuli other than loss of contact inhibition and ROCK-inhibitor presence. Further exploration of the differences between these cell types may assist in optimizing regenerative treatment options for endothelial diseases.

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Section: Discussionmentioning

confidence: 99%

Early and late-onset cell migration from peripheral corneal endothelium

et al. 2023

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“…The corneal endothelium, originating from the neural crest, exhibits cell cycle arrest predominantly in the G0/G1 phase. Therefore, overcoming cell cycle arrest, achieving extensive in vitro expansion, and maintaining a homogeneous and functional population of CECs represent the most challenges in the expansion of adult human CECs [58][59][60][61]. During the expansion and cultivation process, multiple factors, including cell cycle arrest, tight junction disruption, and alterations in cell polarity can lead to cellular senescence and endothelial mesenchymal transition (EnMT) [60,62].…”

Section: In Vitro Culture Strategymentioning

confidence: 99%

Research progress of corneal endothelial cell regeneration and replacement

Li,

Duan,

Zhou

2024

Eye & ENT Research

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Corneal endothelial cells (CECs) are crucial for the maintenance of corneal transparency and normal visual function. Corneal endothelial dysfunction can lead to corneal edema, opacity, and even blindness. Due to the limited proliferative capacity of human CECs and the global shortage of donor cornea, corneal endothelial regeneration and replacement always represent the most challenge in the basic research and clinical treatment of corneal diseases. Although there is a potential existence of corneal endothelial progenitor cells, the efficiency of Descemet stripping without endothelial keratoplasty remains controversial. In recent years, significant advancements have been made in the field of cultured endothelial cell regeneration and artificial material replacement. Here, we reviewed the current research and clinical progress of corneal endothelial cell regeneration and replacement, including the in vitro cultivation of primary human CECs, in vitro differentiation of stem cell‐derived CECs, tissue‐engineered corneal endothelium, and fabrication of artificial corneal endothelium. We also discussed the remaining questions regarding innovating clinical preventive and therapeutic strategies for corneal endothelial dysfunction.

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“…Although these regulations are necessary to maintain quality assurance and patient safety, attaining appropriate GMP licensing can be complex and costly. Standardized methods of cell extraction, isolation, and culture will need to be established and optimized, and laboratory personnel must be trained accordingly [47,48]. The use of xenogeneic products during the cell culture process confers a potential risk of disease transmission, and there are additional regulatory requirements when using these products [49,50].…”

Section: Translation Of Cell-based Research Into Clinical Therapymentioning

confidence: 99%

The role of eye banking with cell-based therapies

Tran

2023

Current Opinion in Ophthalmology

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Purpose of reviewCell-based therapies are an exciting new frontier in managing corneal diseases. The introduction of these novel therapies may provide new alternatives to corneal transplantation and decrease the dependence on donor corneal tissue. These changes have the potential to significantly impact eye banking in the future.Recent findingsThe current article reviews current research involving cell-based therapy for treating corneal disorders, including cultivated limbal stem cell transplantation, limbal mesenchymal stem cells for stromal regeneration, and the use of human-cultivated endothelial cells. We will look at barriers to the development and implementation of these therapies.SummaryAs corneal surgery expands to include cell-based therapies; eye banks will need to redefine their role to support the everchanging landscape of corneal surgery and the decreased demand for corneal donor tissue.

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Challenges in Corneal Endothelial Cell Culture

Cited by 21 publications

References 113 publications

Early and late-onset cell migration from peripheral corneal endothelium

Early and late-onset cell migration from peripheral corneal endothelium

Research progress of corneal endothelial cell regeneration and replacement

The role of eye banking with cell-based therapies

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