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

test_moissonnage_selune
type: Agent

Application of the immunolocalization for researching the human corneal endothelial stem cells
type: Dataset
contact point: Organization
  fn: CCSD
keyword: immunohistochimie
keyword: sdvmheplife-sciences-q-biohuman-health-and-pathology
keyword: therapie-cellulaire-et-genique
keyword: cellule-souche
keyword: cornee-humaine
keyword: immunocytochemistry
keyword: human-cornea
keyword: corneal-endothelial-cells
keyword: immunohistochemistry
keyword: theses
keyword: cellule-epitheliale-corneenne
keyword: cell-and-gene-therapy
keyword: corneal-epithelial-cells
keyword: cellule-endotheliale-corneenne
keyword: infoeu-reposemanticsdoctoralthesis
keyword: stem-cells
keyword: immunocytochimie
keyword: electrotransfer
keyword: electroporation
landing page: https://theses.hal.science/tel-00987968
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…
identifier: NNT: 2011STET011T
issued: 2026-05-05T11:57:17.135258
language: fr
modified: 2026-05-05T11:57:17.135262