Evidence map›Paper›PMID 42831589›Full record

ArticleInvestigative ophthalmology & visual science2026

Divergence Between Marker Expression and Phenotype During Human Corneal Endothelial Cell Expansion.

Filippo Bonelli, Tom Bailey, Iveta Dzivite, Lieve Temmerman, Mor Dickman, Vanessa LaPointe

Abstract readComparative Study
In one paragraph

Article in Investigative ophthalmology & visual science, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
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1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Filippo BonelliDepartment of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht, The Netherlands.
Tom BaileyDepartment of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht, The Netherlands.
Iveta DziviteDepartment of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht, The Netherlands.
Lieve TemmermanDepartment of Pathology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University Medical Center (MUMC+), The Netherlands.
Mor DickmanDepartment of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht, The Netherlands.
Vanessa LaPointeDepartment of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht, The Netherlands.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Purpose: The purpose of this study was to compare whether conventional marker-based assessments adequately reflect phenotypic stability during serial passaging of two established culture protocols for primary human corneal endothelial cells (CEnCs). Methods: Primary CEnCs were isolated from paired donor corneas from 25 individuals (18-44 years of age), and were cultured using 2 established methods. Cells were evaluated at passages 0 to 2 using quantitative morphology (cell area, form factor, nucleus-to-cytoplasm ratio, and cell density), RT-qPCR of endothelial-associated genes, flow cytometry, immunofluorescence, and ELISA quantification of secreted thrombospondin-2 (TSP2). Statistical comparisons were performed using 1- or 2-way ANOVA with multiple comparisons (P < 0.05). Results: CEnCs cultured with method 1 maintained higher cell density, smaller cell area, and greater hexagonality across passages compared with method 2 (P < 0.01 to P < 0.0001). In contrast, transcript levels of selected endothelial-associated markers (ATP1A1 and TJP1) and ZO-1 protein expression were higher in method 2 at later passages (P < 0.01), despite progressive morphological deterioration and loss of membrane localization. Proteins associated with loss of the CEnC phenotype (CD44 and CD10) were significantly elevated in method 2 (P < 0.05 to P < 0.0001), and TSP2 secretion was increased at passage 2 (P < 0.001). Across both methods, most canonical endothelial genes were ≥75% downregulated compared with native tissue. Conclusions: Culture-induced phenotypic drift can occur in CEnCs despite preserved canonical endothelial marker expression. Divergence among marker expression, morphology, localization, and endothelial-to-mesenchymal transition (EndMT) readouts highlights the limits of marker-based evaluation alone. Integrated, multidimensional phenotyping should guide assessment of cultured CEnCs and optimization of expansion protocols, although functional validation is needed to link these readouts to endothelial performance.

Indexed as

BiomarkersEndothelium, CornealGene Expression RegulationAdolescentAdultCell CountCell Culture TechniquesCell ProliferationCells, CulturedEnzyme-Linked Immunosorbent AssayFemaleFlow CytometryHumansMalePhenotypeThrombospondin 2BiomarkersThrombospondin 2ThrombospondinsZonula Occludens-1 Protein

Identifiers

PMID42831589
PMCPMC13657126

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.