Evidence map›Paper›PMID 41123241›Full record

ArticleTranslational vision science & technology2025

Optimized Protocol for Isolation and Culture of Primary Human Corneal Epithelial Cells.

Rongshan Yan, Feeling Y Chen, Ethan S Lindgren, Qi Gao, Yien-Ming Kuo, Danielle M Robertson, Onur Cil, Matilda F Chan, Neel D Pasricha

Abstract read
In one paragraph

Article in Translational vision science & technology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

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

1 citing paper in PubMed.

  1. Review
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.

Rongshan YanDepartment of Ophthalmology, University of California San Francisco, San Francisco, CA, USA.
Feeling Y ChenDepartment of Cell & Tissue Biology, University of California San Francisco, San Francisco, CA, USA.
Ethan S LindgrenDepartment of Ophthalmology, University of California San Francisco, San Francisco, CA, USA.
Qi GaoDepartment of Cardiovascular Disease, The first Hospital of Jilin University, Changchun, China.
Yien-Ming KuoDepartment of Ophthalmology, University of California San Francisco, San Francisco, CA, USA.
Danielle M RobertsonDepartment of Ophthalmology, UT Southwestern Medical Center, Dallas, TX, USA.
Onur CilDepartment of Pediatrics, University of California San Francisco, San Francisco, CA, USA.
Matilda F ChanDepartment of Ophthalmology, University of California San Francisco, San Francisco, CA, USA.
Neel D PasrichaDepartment of Ophthalmology, University of California San Francisco, San Francisco, CA, USA.

Funding

Rapid-Prototyping and Design CoreP30EY002162 · NEI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI ULLIAN, ERIK M · 1985 to 2024
$16.8M
Vision Bioinformatics and ComputationP30EY037668 · NEI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI Erik M Ullian · 2025 to 2026
$2.0M
SLC26A3 (DRA) Inhibitors for Treatment of Hyperoxaluria and NephrolithiasisR01DK126070 · NIDDK · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI CIL, ONUR · 2020 to 2024
$1.6M
Roles of CaSR in ocular surface health and diseaseR01EY036139 · NEI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI Onur Cil · 2024 to 2026
$1.2M
Human Ocular Surface ElectrophysiologyK08EY033859 · NEI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI Neel Dave Pasricha · 2023 to 2026
$877k
NEI NIH HHS K08 EY033859NEI NIH HHS P30 EY002162NEI NIH HHS P30 EY037668NEI NIH HHS R01 EY036139NIDDK NIH HHS R01 DK126070
6 · The paper itself

Abstract

Purpose: To establish a reliable method for isolating and culturing high-purity primary human corneal epithelial cells (HCECs) for ophthalmic drug testing. Methods: We present a detailed, step-by-step protocol for the efficient isolation and culture of primary HCECs. This protocol includes the characterization of HCEC morphological responses to varying Ca2+ concentrations in the culture medium. Additionally, immunofluorescence staining with well-established markers is used to identify and confirm the cell types in vitro. A Ca2+ assay is performed to validate the functionality of the cultured primary HCECs. Results: By following the procedures detailed in this protocol, high-purity primary HCECs with strong proliferative capacity and preserved morphological integrity are obtained. Immunofluorescence staining confirms the presence of both limbal stem cells and differentiated corneal epithelial cells in vitro. Additionally, the functional assay demonstrates that the cultured primary HCECs retain the ability to respond to external Ca²⁺ stimuli. Conclusions: This optimized protocol enhances the efficiency and reliability of primary HCECs isolation and culture, enabling the development of a robust in vitro model for studying the mechanisms of ocular diseases. Translational Relevance: Successfully cultured primary HCECs in vitro bridges the gap between laboratory findings and clinical applications, facilitating advancements in therapeutic development.

Indexed as

Cell Culture TechniquesCell SeparationEpithelial CellsEpithelium, CornealCalciumCell DifferentiationCell ProliferationCells, CulturedHumansCalcium

Identifiers

PMID41123241
PMCPMC12553468

What Socratic holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

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 Socratic graph.