ISSN Print 2500–1094
ISSN Online 2542–1204
BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)
Copyright: © 2025 by the authors. Licensee: Pirogov University.
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (CC BY).
This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (CC BY).
ORIGINAL RESEARCH
Differentiation of iPSCs into corneal epithelial precursors in three-dimensional In vitro culture
About authors
1 Pirogov Russian National Research Medical University, Moscow, Russia
2 Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia
Correspondence should be addressed: Nadezhda G. Gurskaya
Ostrovityanova, 1, str. 1, Moscow, 117997, Russia; ur.liam@ayaksrugn
About paper
Funding: the work was supported by a grant from the Ministry of Health of the Russian Federation, project No. 124021000001-09.
Acknowledgements: the authors thank M. A. Lagarkova, Corresponding Member of the Russian Academy of Sciences, Professor, for fruitful discussion of the work and critical comments.
Author contribution: Zhigmitova EB — design and conduct of the experiment, Kosykh AV — material processing, micrography, preparation of the article, Gurskaya NG — concept, design of the work, primary manuscript editing.
Received: 2025-11-25
Accepted: 2025-12-12
Published online: 2025-12-27
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Fig. 1. Protocol for generating EBs and their subsequent differentiation into corneal epithelial cells. D(–12) — the beginning of the experiment. D(1) — the first day of the differentiation protocol. D(22) — the last day of differentiation; collection of organoids for subsequent analysis
Fig. 2. Darkfield photo of organoids on the 22nd day of differentiation. There are both translucent spheres and pigmented areas. * — translucent zones at the periphery of the organoid, presumably corresponding to corneal regions; ** — pigmented areas of the organoid, presumably corresponding to retinal pigment epithelium (RPE) regions. Each scale division is 1 mm
Fig. 3. Immunohistochemical staining of cryopresections of organoids on the 22nd day of differentiation. Reaction with antibodies to keratin 3 (red) and reaction with antibodies to type VII collagen (green). The cores were additionally colored with DAPI. The scale segment is 50 μm
