Early and late-onset cell migration from peripheral corneal endothelium

Abstract

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.

A Case of Transparent Cornea Maintained after Removal of Dislocated Graft from Descemet’s Stripping Automated Endothelial Keratoplasty

“…Corneal endothelial cells migrate both in vitro and in vivo [7][8][9], and migration from the graft to the host cornea has been reported [7]. In this case, endothelial cells from the corneal graft possibly migrated to the host cornea, which may have restored the corneal endothelial pump function and maintained corneal transparency even after graft removal.…”

Transcription factor 4 promotes increased corneal endothelial cellular migration by altering microtubules in Fuchs endothelial corneal dystrophy

“…This regenerative wound healing response is stimulated with DSO, and functional recovery is based on CEC migration from the periphery to the central cornea 18 – 21 . Recently, two distinct migrating cell populations that display collective migration from the peripheral to central endothelium have been observed in outer corneal graft rims and a thermo-reversible hydrogel 9 . These two morphologically distinct populations were identified to be an early-onset migrating population triggered by disrupted contact inhibition and another late-onset population showing higher proliferative capacity and appearing less differentiated 9 .…”

“…Recently, two distinct migrating cell populations that display collective migration from the peripheral to central endothelium have been observed in outer corneal graft rims and a thermo-reversible hydrogel 9 . These two morphologically distinct populations were identified to be an early-onset migrating population triggered by disrupted contact inhibition and another late-onset population showing higher proliferative capacity and appearing less differentiated 9 . One possibility is that the peripheral corneal endothelium acts as a reserve to regenerate the corneal endothelium after injury or degeneration through these two populations, where the late-onset population may be derived from the progenitor rich transition zone 9 , 64 – 70 .…”

“…These two morphologically distinct populations were identified to be an early-onset migrating population triggered by disrupted contact inhibition and another late-onset population showing higher proliferative capacity and appearing less differentiated 9 . One possibility is that the peripheral corneal endothelium acts as a reserve to regenerate the corneal endothelium after injury or degeneration through these two populations, where the late-onset population may be derived from the progenitor rich transition zone 9 , 64 – 70 . Whether this occurs in vivo and if this migration pattern is altered in FECD remains to be investigated.…”

Squishy matters – Corneal mechanobiology in health and disease

“…In a variety of corneal endothelial cell culture models, ROCK inhibition increases cell migration, proliferation, and adhesion while inhibiting apoptosis in vitro ( Jung et al, 2021 ; Kim et al, 2022 ; Lee et al, 2016 ; Miron et al, 2023 ; Okumura et al, 2009 ; Peh et al, 2015 ; Toda et al, 2017 ). For example, Okumura and colleagues demonstrated Y27632 increased cell adhesion and proliferation and decreased apoptosis in cultured cynomolgus macaque endothelial cells ( Okumura et al, 2009 ).…”