Application of the immunolocalization for researching the human corneal endothelial stem cells
type: Dataset
description: The control of corneal transparency depends on the integrity of the mono-stratified corneal endothelium, which is considered devoid of regenerative capacity after birth in humans. In pathological conditions leading to blindness by irreversible corneal edema, the significant losses of endothelial cells (ECs) are not replaced efficiently, indicating that neither new ECs derived from stem cells (SC) nor the division of ECs neighboring the lesions can contribute to a form of regeneration. However, several works of the last 25 years demonstrated that ECs possess residual capacity of proliferation ex vivo, and more recently, two teams suggested the existence of SC or endothelial progenitors located in the corneal periphery. In this thesis work, we have firstly optimized an immunostaining technique specially adapted to intact endothelium of flat mounted whole corneas. Consequently, we now have simple protocols of fixation at the right temperature, and of antigen retrieval that allow detecting multiple proteins with a clear subcellular localization. Using important series of non-stored and of organ cultured corneas, and thanks to this technique, we investigated the cell cycle status of ECs and the location of potential SC in human corneal endothelium. Our results indicate that ECs homogeneously express positive regulators (PCNA, MCM2, cyclin D1, cyclin E and cyclin A) as well as negative regulators of the cell cycle (P16, P27); several original descriptions have been made: diffuse cytoplasmic location of MCM2, paranuclear location of cyclin D1, absence of P21. The expression patterns suggest that ECS could be arrested after the restriction point of the G1 phase and that numerous mechanism of DNA repair are stimulated in ECs exposed to an important oxidative stress throughout live. We identified a novel organization of the micro-anatomy of the endothelial periphery and extreme-periphery, where cells accumulate in multiple small multilayered clusters connected radial centripe…
landing page: https://theses.hal.science/tel-00987968
identifier: NNT: 2011STET011T
keyword: cell-and-gene-therapy
keyword: cellule-endotheliale-corneenne
keyword: cellule-epitheliale-corneenne
keyword: cellule-souche
keyword: corneal-endothelial-cells
keyword: corneal-epithelial-cells
keyword: cornee-humaine
keyword: electroporation
keyword: electrotransfer
keyword: human-cornea
keyword: immunocytochemistry
keyword: immunocytochimie
keyword: immunohistochemistry
keyword: immunohistochimie
keyword: infoeu-reposemanticsdoctoralthesis
keyword: sdvmheplife-sciences-q-biohuman-health-and-pathology
keyword: stem-cells
keyword: therapie-cellulaire-et-genique
keyword: theses
issued: 2026-05-05T11:57:17.135258
modified: 2026-05-05T11:57:17.135262
language: fr
contact point: Organization
  fn: CCSD

test_moissonnage_selune
type: Agent

Application of the immunolocalization for researching the human corneal endothelial stem cells
type: Distribution
format: HTML
access URL: https://theses.hal.science/tel-00987968
issued: 2026-05-05T11:57:17.137353
modified: 2026-05-05T11:57:17.045270