Successful culture of human transition zone cells

Zhang, Jie; Ahmad, Amatul Mateen; Ng, Hannah; Shi, Jane; McGhee, Charles N J; Patel, Dipika V.

doi:10.1111/ceo.13756

Cited by 10 publications

(7 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, Zhang et al reported endothelial cell regeneration from human "wider TZ" (~2 mm width), comprising of Schwalbe's line, a portion of PE and TM, by explant culture. They showed the proliferation of endothelial-like cells with the expression of ZO-1, Na + / K + -ATPase, and Col8A2 [55]. However, the source of cultured cells from TZ or PE was not determined.…”

Section: Ex Vivo Propagation Of Tz Cells and Generation Of The Corneamentioning

confidence: 99%

Regenerative capacity of the corneal transition zone for endothelial cell therapy

et al. 2020

View full text Add to dashboard Cite

The corneal endothelium located on the posterior corneal surface is responsible for regulating stromal hydration. This is contributed by a monolayer of corneal endothelial cells (CECs), which are metabolically active in a continuous fluid-coupled efflux of ions from the corneal stroma into the aqueous humor, preventing stromal over-hydration and preserving the orderly arrangement of stromal collagen fibrils, which is essential for corneal transparency. Mature CECs do not have regenerative capacity and cell loss due to aging and diseases results in irreversible stromal edema and a loss of corneal clarity. The current gold standard of treatment for this worldwide blindness caused by corneal endothelial failure is the corneal transplantation using cadaveric donor corneas. The top indication is Fuchs corneal endothelial dystrophy/degeneration, which represents 39% of all corneal transplants performed. However, the global shortage of transplantable donor corneas has restricted the treatment outcomes, hence instigating a need to research for alternative therapies. One such avenue is the CEC regeneration from endothelial progenitors, which have been identified in the peripheral endothelium and the adjacent transition zone. This review examines the evidence supporting the existence of endothelial progenitors in the posterior limbus and summarizes the existing knowledge on the microanatomy of the transitional zone. We give an overview of the isolation and ex vivo propagation of human endothelial progenitors in the transition zone, and their growth and differentiation capacity to the corneal endothelium. Transplanting these bioengineered constructs into in vivo models of corneal endothelial degeneration will prove the efficacy and viability, and the long-term maintenance of functional endothelium. This will develop a novel regenerative therapy for the management of corneal endothelial diseases.

show abstract

Section: Ex Vivo Propagation Of Tz Cells and Generation Of The Corneamentioning

confidence: 99%

Regenerative capacity of the corneal transition zone for endothelial cell therapy

et al. 2020

View full text Add to dashboard Cite

show abstract

“…When spheres were injected into the anterior chamber of rabbit eyes with corneal deficiency, they formed a functional endothelium [ 30 , 31 ]. Zhang et al demonstrated that TZ cells can proliferate and differentiate into CECs by culturing TZ cells from donor corneal rims [ 32 ]. The TZ cells grew from the explant after 20 days of culture, exhibiting initially a rounder polygonal morphology, that become gradually more elongated and fibroblastic during passaging and finally polygonal 2 to 3 passages before senescence.…”

Section: Discussionmentioning

confidence: 99%

“…Although endothelial-to-mesenchymal transition (EMT) has been described to occur when CECs are cultured over multiple passages and cells adopted a more elongated morphology [ 33 ], highly proliferative TZ cells first adopt a fibroblast-like morphology and later transform to a polygonal morphology [ 32 ]. Previous studies also reported that there is no association of the TZ dimension or proliferative characteristics with donor age, ethnicity, or cell density which is in agreement with our findings that the outer rims showing the late-onset cells were isolated from old donors (68–80-years-old) with variable ECD (1800–2400 cell/mm 2 ), and preservation time before culture (8–26 days) [ 27 ].…”

Section: Discussionmentioning

confidence: 99%

Early and late-onset cell migration from peripheral corneal endothelium

et al. 2023

View full text Add to dashboard Cite

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.

show abstract

“…However, there are very few reports of immunostaining of corneal endothelial cells in dogs. Na + /K + ‐ATPase and ZO‐1 are representative markers of corneal endothelial cells in humans 14,23–26 …”

Section: Discussionmentioning

confidence: 99%

Primary cell culture of canine corneal endothelial cells

Tajima

Okada

Kudo

et al. 2021

Veterinary Ophthalmology

View full text Add to dashboard Cite

Objective To establish a primary cell culture and clarify the characteristics of canine corneal endothelial cells in vitro. Procedures The eyes were enucleated from dogs that were euthanized for reasons unrelated to this study. Enucleated canine eyes were dissected, and the intact corneas were isolated from the globes. Using enzymes, the corneal endothelial cells were dispersed from the cornea. The obtained canine corneal endothelial cells were cultured in a cell culture dish. Cultured corneal endothelial cells were morphologically evaluated using phase‐contrast microscopy. Immunohistochemical analysis of the cultured cells, particularly of the corneal endothelial cell marker, zonula occludens‐1 (ZO‐1), Na+/K+‐ATPase, and vimentin, was performed to clarify whether the cultured cells were actually corneal endothelial cells. Furthermore, the post‐passage morphology of cultured cells was evaluated. Results Canine primary cultured corneal endothelial cells showed morphologically small, cobblestone‐like structures. The isolated cells had proliferative ability in vitro and demonstrated positive expression of the corneal endothelial cell markers, ZO‐1, Na+/K+‐ATPase, and vimentin. However, repeated passages resulted in larger cell sizes as assessed by phase‐contrast microscopy. Repeated passages also resulted in lower cell density. Conclusions This study demonstrated the successful culture of canine corneal endothelial cells. This might enhance the understanding of corneal endothelial cell characteristics in dogs.

show abstract

Successful culture of human transition zone cells

Cited by 10 publications

References 31 publications

Regenerative capacity of the corneal transition zone for endothelial cell therapy

Regenerative capacity of the corneal transition zone for endothelial cell therapy

Early and late-onset cell migration from peripheral corneal endothelium

Primary cell culture of canine corneal endothelial cells

Contact Info

Product

Resources

About