Sofija Andjelić1, Kazimir Drašlar2, Xhevat Lumi1, Xiaohe Yan3, Joachim Graw3, Andrea Facskó4, Marko Hawlina1, Goran Petrovski1,4,5. 1. Eye Hospital, University Medical Centre, Ljubljana, Slovenia. 2. Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia. 3. Helmholtz Center Munich German Research Center for Environmental Health, Institute of Developmental Genetics, Neuherberg, Germany. 4. Department of Ophthalmology, Faculty of Medicine, University of Szeged, Szeged, Hungary. 5. Stem Cells and Eye Research Laboratory, Department of Biochemistry and Molecular Biology, and Apoptosis and Genomics Research Group of the Hungarian Academy of Sciences, University of Debrecen, Debrecen, Hungary.
Abstract
PURPOSE: To determine the structural characteristics of lens epithelial cells (LECs) found on the anterior portion of the lens capsule and their pluripotency, proliferating and migrating potential when grown ex vivo with relevance to posterior capsular opacification (PCO) after cataract surgery. METHODS: The explants of anterior portion of the lens capsule consisting of monolayer of LECs were obtained from uneventful cataract surgery and were cultivated under adherent conditions. The size and shape of the outgrowing cells were recorded by scanning electron microscopy (SEM), while their migration and proliferation potential were followed using light microscopy. Positivity for proliferation (Ki-67)- and pluripotency (Sox2)-specific markers were tested by immunofluorescent staining. RESULTS: The proliferation and migration of anterior portion of the lens capsule's LECs filling up the denuded and reverse side regions of the lens capsule as well as their growth on glass culture surfaces could be followed by light microscopy and SEM, while the distribution of LECs and their morphology could be analysed in detail by SEM. The expression of Ki-67 and Sox2 in LECs growing adherently on human anterior portion of the lens capsule could also be detected. CONCLUSIONS: Classic light microscopy and SEM can be used to show that human anterior portion of the lens capsule harbours LECs that can proliferate and migrate, suggesting their pluripotency or putative stem cell nature. Similarly, morphological techniques can be used to study PCO and the effect different drugs or physical treatments have against PCO development.
PURPOSE: To determine the structural characteristics of lens epithelial cells (LECs) found on the anterior portion of the lens capsule and their pluripotency, proliferating and migrating potential when grown ex vivo with relevance to posterior capsular opacification (PCO) after cataract surgery. METHODS: The explants of anterior portion of the lens capsule consisting of monolayer of LECs were obtained from uneventful cataract surgery and were cultivated under adherent conditions. The size and shape of the outgrowing cells were recorded by scanning electron microscopy (SEM), while their migration and proliferation potential were followed using light microscopy. Positivity for proliferation (Ki-67)- and pluripotency (Sox2)-specific markers were tested by immunofluorescent staining. RESULTS: The proliferation and migration of anterior portion of the lens capsule's LECs filling up the denuded and reverse side regions of the lens capsule as well as their growth on glass culture surfaces could be followed by light microscopy and SEM, while the distribution of LECs and their morphology could be analysed in detail by SEM. The expression of Ki-67 and Sox2 in LECs growing adherently on human anterior portion of the lens capsule could also be detected. CONCLUSIONS: Classic light microscopy and SEM can be used to show that human anterior portion of the lens capsule harbours LECs that can proliferate and migrate, suggesting their pluripotency or putative stem cell nature. Similarly, morphological techniques can be used to study PCO and the effect different drugs or physical treatments have against PCO development.
Authors: Jovana Bisevac; Natalia S Anisimova; Richárd Nagymihály; Olav Kristianslund; Kirankumar Katta; Agate Noer; Ilias H Sharafetdinov; Liv Drolsum; Morten C Moe; Boris E Malyugin; Goran Petrovski Journal: Sci Rep Date: 2020-11-25 Impact factor: 4.379