Cell Pattern in Adult Human Corneal Endothelium

Wörner, C. H.; Olguín, A.; Ruíz-García, José L.; Garzón, Nuria

doi:10.1371/journal.pone.0019483

Cited by 31 publications

(25 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Cells of the healthy human corneal endothelium are generally hexagonal in shape (60,62). Hence, an important morphological characteristic of hCECs expanded in vitro is the maintenance of their polygonal/hexagonal cellular shape in culture.…”

Section: Morphometry Analysis Of Cellular Circularitymentioning

confidence: 99%

Propagation of Human Corneal Endothelial Cells: A Novel Dual Media Approach

et al. 2015

View full text Add to dashboard Cite

Corneal endothelium-associated corneal blindness is the most common indication for corneal transplantation. Restorative corneal transplant surgery is the only option to reverse the blindness, but a global shortage of donor material remains an issue. There are immense clinical interests in the development of alternative treatment strategies to alleviate current reliance on donor materials. For such endeavors, ex vivo propagation of human corneal endothelial cells (hCECs) is required, but current methodology lacks consistency, with expanded hCECs losing cellular morphology to a mesenchymal-like transformation. In this study, we describe a novel dual media culture approach for the in vitro expansion of primary hCECs. Initial characterization included analysis of growth dynamics of hCECs grown in either proliferative (M4) or maintenance (M5) medium. Subsequent comparisons were performed on isolated hCECs cultured in M4 alone against cells expanded using the dual media approach. Further characterizations were performed using immunocytochemistry, quantitative real-time PCR, and gene expression microarray. At the third passage, results showed that hCECs propagated using the dual media approach were homogeneous in appearance, retained their unique polygonal cellular morphology, and expressed higher levels of corneal endothelium-associated markers in comparison to hCECs cultured in M4 alone, which were heterogeneous and fibroblastic in appearance. Finally, for hCECs cultured using the dual media approach, global gene expression and pathway analysis between confluent hCECs before and after 7-day exposure to M5 exhibited differential gene expression associated predominately with cell proliferation and wound healing. These findings showed that the propagation of primary hCECs using the novel dual media approach presented in this study is a consistent method to obtain bona fide hCECs. This, in turn, will elicit greater confidence in facilitating downstream development of alternative corneal endothelium replacement using tissue-engineered graft materials or cell injection therapy.

show abstract

Section: Morphometry Analysis Of Cellular Circularitymentioning

confidence: 99%

Propagation of Human Corneal Endothelial Cells: A Novel Dual Media Approach

et al. 2015

View full text Add to dashboard Cite

show abstract

“…In the adult, the average cell density is B3000 cells/mm 2 and the percentage of hexagonal cells is about 75%. 26,27 The density of corneal endothelial cells and their surface changes throughout life are noted in Figure 3. From the second to the eighth decade, the cell density declines to about 2600 cells/mm 2 and the percentage of hexagonal cells decreases to B60%.…”

Section: Corneal Endothelium Physiologymentioning

confidence: 99%

Corneal endothelium: developmental strategies for regeneration

Zavala

Jaime

Barrientos

et al. 2013

Eye

113

View full text Add to dashboard Cite

The main treatment available for restoration of the corneal endothelium is keratoplasty. This procedure is faced with several difficulties, including the shortage of donor tissue, postsurgical complications associated with the use of drugs to prevent immune rejection, and a significant increase in the occurrence of glaucoma. Recently, surgical procedures such as Descemet's stripping endothelial keratoplasty have focused on the transplant of corneal endothelium, yielding better visual results but still facing the need for donor tissue. The emergent strategies in the field of cell biology and tissue cultivation of corneal endothelial cells aim at the production of transplantable endothelial cell sheets. Cell therapy focuses on the culture of corneal endothelial cells retrieved from the donor, in the donor's cornea, followed by transplantation into the recipient. Recently, research has focused on overcoming the challenge of harvesting human corneal endothelial cells and the generation of new biomembranes to be used as cell scaffolds in surgical procedures. The use of corneal endothelial precursors from the peripheral cornea has also demonstrated to be effective and represents a valuable tool for reducing the risk of rejection in allogeneic transplants. Several animal model reports also support the use of adult stem cells as therapy for corneal diseases. Current results represent important progresses in the development of new strategies based on alternative sources of tissue for the treatment of corneal endotheliopathies. Different databases were used to search literature: PubMed, Google Books, MD Consult, Google Scholar, Gene Cards, and NCBI Books. The main search terms used were: 'cornea AND embryology AND transcription factors', 'human endothelial keratoplasty AND risk factors', '(cornea OR corneal) AND (endothelium OR endothelial) AND cell culture', 'mesenchymal stem cells AND cell therapy', 'mesenchymal stem cells AND cornea', and 'stem cells AND (cornea OR corneal) AND (endothelial OR endothelium)'.

show abstract

“…Mature CECs are metabolically active, with continuous ATPase activity for the fluid-coupled active transport of ions from the corneal stroma to aqueous humor [2]. This “leaky barrier and ionic pump” activity regulates the stromal hydration and prevents edema in maintaining the corneal transparency, which is necessary for normal vision [3,4,5]. Under optical coherence tomography, it is evident that the CE terminates at the Schwalbe’s line in the corneal periphery [6].…”

Section: Introductionmentioning

confidence: 99%

Characterization of Human Transition Zone Reveals a Putative Progenitor-Enriched Niche of Corneal Endothelium

Yam

Seah

Yusoff

et al. 2019

Cells

View full text Add to dashboard Cite

The corneal endothelium regulates corneal hydration to maintain the transparency of cornea. Lacking regenerative capacity, corneal endothelial cell loss due to aging and diseases can lead to corneal edema and vision loss. There is limited information on the existence of corneal endothelial progenitors. We conducted ultrastructural examinations and expression analyses on the human transition zone (TZ) at the posterior limbus of corneal periphery, to elucidate if the TZ harbored progenitor-like cells, and to reveal their niche characteristics. Within the narrow TZ (~190 μm width), the inner TZ—adjacent to the peripheral endothelium (PE)—contained cells expressing stem/progenitor markers (Sox2, Lgr5, CD34, Pitx2, telomerase). They were located on the inner TZ surface and in its underlying stroma. Lgr5 positive cells projected as multicellular clusters into the PE. Under transmission electron microscopy and serial block face-scanning electron microscopy and three-dimensional (3D) reconstruction, the terminal margin of Descemet’s membrane was inserted beneath the TZ surface, with the distance akin to the inner TZ breadth. Porcine TZ cells were isolated and proliferated into a confluent monolayer and differentiated to cells expressing corneal endothelial markers (ZO1, Na+K+ATPase) on cell surface. In conclusion, we have identified a novel inner TZ containing progenitor-like cells, which could serve the regenerative potential for corneal endothelium.

show abstract

Cell Pattern in Adult Human Corneal Endothelium

Cited by 31 publications

References 39 publications

Propagation of Human Corneal Endothelial Cells: A Novel Dual Media Approach

Propagation of Human Corneal Endothelial Cells: A Novel Dual Media Approach

Corneal endothelium: developmental strategies for regeneration

Characterization of Human Transition Zone Reveals a Putative Progenitor-Enriched Niche of Corneal Endothelium

Contact Info

Product

Resources

About