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Peer-reviewedCC BYScientific Reports

Optimization of culture conditions for the human corneal epithelial cell line IM-HCEpiC and evaluation of its extracellular vesicles in repair

  • Sara Galindo1
  • Estefanía González-Martín2
  • Helen Gutiérrez2
  • Yolanda Diebold3
  • Laura García-Posadas2
  1. 1Ocular Surface Group, Instituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de Valladolid
  2. 2Nanotechnological Solutions for Ocular Functional Recovery Group, Instituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de Valladolid
  3. 3Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Instituto de Salud Carlos III
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Abstract

Corneal epithelial defects are a major cause of vision loss, highlighting the need for reliable in vitro models to study ocular surface pathophysiology and for therapeutic development. In this study, we establish a simplified and reproducible protocol for culturing the human corneal epithelial cell line IM-HCEpiC under standardized culture conditions, avoiding coated flasks and proprietary media of unknown composition. We compared proliferation rates across different seeding densities and culture media formulations, identifying DMEM/F12 supplemented with fetal bovine serum as the optimal compositionally defined medium, while collagen coating does not significantly improve growth. Under these conditions, IM-HCEpiC cells expressed corneal epithelial markers (CK3, CK12, ZO-1, E-cadherin, and Pax-6) and displayed the expected response to oxidative stress and antioxidant treatment, supporting the preservation of key epithelial characteristics. Using this optimized culture system, extracellular vesicles (EVs) were isolated from IM-HCEpiC secretomes, characterized, and functionally evaluated. IM-HCEpiC-derived EVs promote corneal wound closure and increased Ki67 expression. Therefore, this study establishes a simplified, cost-effective, and reproducible culture protocol for IM-HCEpiC cells while preserving key corneal epithelial characteristics, supporting their use in corneal epithelial biology and EV-based research.

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Cite

BibTeXRIS
BibTeX
@article{galindo2026optimization,
  title = {{Optimization of culture conditions for the human corneal epithelial cell line IM-HCEpiC and evaluation of its extracellular vesicles in repair}},
  author = {Sara Galindo and Estefanía González-Martín and Helen Gutiérrez and Yolanda Diebold and Laura García-Posadas},
  journal = {Scientific Reports},
  year = {2026},
  month = jul,
  doi = {10.1038/s41598-026-61839-2},
  url = {https://doi.org/10.1038/s41598-026-61839-2},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Optimization of culture conditions for the human corneal epithelial cell line IM-HCEpiC and evaluation of its extracellular vesicles in repair
AU  - Sara Galindo
AU  - Estefanía González-Martín
AU  - Helen Gutiérrez
AU  - Yolanda Diebold
AU  - Laura García-Posadas
PY  - 2026
DA  - 2026/07/01
JO  - Scientific Reports
DO  - 10.1038/s41598-026-61839-2
UR  - https://doi.org/10.1038/s41598-026-61839-2
AB  - Corneal epithelial defects are a major cause of vision loss, highlighting the need for reliable in vitro models to study ocular surface pathophysiology and for therapeutic development. In this study, we establish a simplified and reproducible protocol for culturing the human corneal epithelial cell line IM-HCEpiC under standardized culture conditions, avoiding coated flasks and proprietary media of unknown composition. We compared proliferation rates across different seeding densities and culture media formulations, identifying DMEM/F12 supplemented with fetal bovine serum as the optimal compositionally defined medium, while collagen coating does not significantly improve growth. Under these conditions, IM-HCEpiC cells expressed corneal epithelial markers (CK3, CK12, ZO-1, E-cadherin, and Pax-6) and displayed the expected response to oxidative stress and antioxidant treatment, supporting the preservation of key epithelial characteristics. Using this optimized culture system, extracellular vesicles (EVs) were isolated from IM-HCEpiC secretomes, characterized, and functionally evaluated. IM-HCEpiC-derived EVs promote corneal wound closure and increased Ki67 expression. Therefore, this study establishes a simplified, cost-effective, and reproducible culture protocol for IM-HCEpiC cells while preserving key corneal epithelial characteristics, supporting their use in corneal epithelial biology and EV-based research.
SN  - 2045-2322
ER  -