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

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

SUBMITTER: Yan R 

PROVIDER: S-EPMC12553468 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature