Evidence map›Paper›PMID 25734654›Full record

ArticlePloS one2015

Serum-free and xenobiotic-free preservation of cultured human limbal epithelial cells.

Oeygunn Utheim, Rakibul Islam, Torstein Lyberg, Borghild Roald, Jon Roger Eidet, Maria Fideliz de la Paz, Darlene A Dartt, Sten Raeder, Tor Paaske Utheim

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In one paragraph

Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers, 1 of them a synthesis that pooled it.

0numbers the graph read from it
0cells of the map it votes in
14citing papers in PubMed, 1 pooled it
–field-weighted citation impact
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

14 citing papers in PubMed, 1 synthesis or guideline pooled it.

  1. Pooled it
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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

9 authors.

Oeygunn UtheimDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, Norway; Department of Ophthalmology, Oslo University Hospital, Oslo, Norway.
Rakibul IslamDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, Norway; Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.
Torstein LybergDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.
Borghild RoaldDepartment of Pathology, Oslo University Hospital, Oslo, Norway.
Jon Roger EidetDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.
Maria Fideliz de la PazInstitut Universitari Barraquer, Universitat Autonoma de Barcelona, Barcelona, Spain.
Darlene A DarttSchepens Eye Research Institute / Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA, United States of America.
Sten RaederThe Norwegian Dry Eye Clinic, Oslo/Tromsoe, Norway.
Tor Paaske UtheimDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, Norway; Department of Ophthalmology, Oslo University Hospital, Oslo, Norway; Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway.

Funding

Conjunctival Goblet Cell Mucin Secretion in Inflammation and Its ResolutionR01EY019470 · NEI · SCHEPENS EYE RESEARCH INSTITUTE · PI DARTT, DARLENE A · 2009 to 2023
$7.4M
NEI NIH HHS R01 EY019470
6 · The paper itself

Abstract

aim/purpose of the studyTo develop a one-week storage method, without serum and xenobiotics, that would maintain cell viability, morphology, and phenotype of cultured human limbal epithelial sheets. MATERIALS AND

methodsHuman limbal explants were cultured on intact human amniotic membranes for two weeks. The sheets were stored in a hermetically sealed container at 23°C in either a serum-free medium with selected animal serum-derived compounds (Quantum 286) or a xenobiotic-free medium (Minimal Essential Medium) for 4 and 7 days. Stored and non-stored cultures were analyzed for cell viability, amniotic membrane and epithelial sheet thickness, and a panel of immunohistochemical markers for immature cells (ΔNp63α, p63, Bmi-1, C/EBP∂, ABCG2 and K19), differentiated cells (K3 and Cx43), proliferation (PCNA), and apoptosis (Caspase-3).

resultsThe cell viability of the cultures was 98 ± 1% and remained high after storage. Mean central thickness of non-stored limbal epithelial sheets was 23 ± 3 μm, and no substantial loss of cells was observed after storage. The non-stored epithelial sheets expressed a predominantly immature phenotype with ΔNp63α positivity of more than 3% in 9 of 13 cultures. After storage, the expression of ABCG2 and C/EBP∂ was reduced for the 7 day Quantum 286-storage group; (P = 0.04), and Bmi-1 was reduced after 4 day Quantum 286-storage; (P = 0.02). No other markers varied significantly. The expression of differentiation markers was unrelated to the thickness of the epithelia and amniotic membrane, apart from ABCG2, which correlated negatively with thickness of limbal epithelia (R = -0.69, P = 0.01) and ΔNp63α, which correlated negatively with amniotic membrane thickness (R = -0.59, P = 0.03).

conclusionLimbal epithelial cells cultured from explants on amniotic membrane can be stored at 23°C in both serum-free and xenobiotic-free media, with sustained cell viability, ultrastructure, and ΔNp63α-positivity after both 4 and 7 days.

Indexed as

AmnionATP-Binding Cassette TransportersATP Binding Cassette Transporter, Subfamily G, Member 2BiomarkersCaspase 3CCAAT-Enhancer-Binding Protein-deltaCell DifferentiationCell SurvivalConnexin 43Epithelial CellsGene ExpressionHumansKeratin-19Limbus CorneaeMembrane ProteinsMitogen-Activated Protein Kinase 7ABCG2 protein, humanATP-Binding Cassette TransportersATP Binding Cassette Transporter, Subfamily G, Member 2BiomarkersCaspase 3CCAAT-Enhancer-Binding Protein-deltaCEBPD protein, humanCKAP4 protein, humanConnexin 43Keratin-19MAPK7 protein, humanMembrane ProteinsMitogen-Activated Protein Kinase 7Neoplasm ProteinsProliferating Cell Nuclear Antigen

Identifiers

PMID25734654
PMCPMC4348416

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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.