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Updated: Dec 7, 2025

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Methods for Investigating Corneal Cell Interactions and Extracellular Vesicles In Vitro.

Tina B McKay1, Xiaoqing Guo1, Audrey E K Hutcheon1

  • 1Schepens Eye Research Institute of Massachusetts Eye and Ear and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.

Current Protocols in Cell Biology
|September 28, 2020
PubMed
Summary
This summary is machine-generated.

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This study details methods for creating advanced in vitro human cornea models using cell culture techniques. These tissue-engineered corneas enable research into drug permeability, scarring, and regeneration.

Area of Science:

  • Biomedical Engineering
  • Ophthalmology Research
  • Tissue Engineering

Background:

  • Increasing shift towards human-centric research models in science and medicine.
  • Development of sophisticated in vitro tissue models using human-derived cells.
  • Need for advanced in vitro models of the human cornea for physiological and disease studies.

Purpose of the Study:

  • To describe methods for cultivating human corneal cells for in vitro applications.
  • To provide protocols for assembling and culturing 3D corneal constructs.
  • To detail the isolation of extracellular vesicles from corneal cell conditioned medium.

Main Methods:

  • Cultivation of human corneal keratocytes, fibroblasts, epithelial, and endothelial cells.
  • Assembly and culturing of 3D corneal stromal constructs.
Keywords:
co-culturecorneal endothelial cellscorneal epithelial cellsextracellular vesicleskeratocytes

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  • Co-culture of epithelial- and endothelial-stromal components.
  • Isolation of extracellular vesicles from conditioned medium.

    • Main Results:

    • Established protocols for isolating and culturing various human corneal cell types.
    • Successful assembly and maintenance of 3D corneal stromal and co-culture constructs.
    • Methods for cryopreservation of corneal cells developed.
    • Protocols for extracellular vesicle isolation from corneal cell cultures provided.

    Conclusions:

    • Tissue-engineered human corneas can be reliably grown and maintained in vitro for extended periods.
    • These models are suitable for studying extracellular matrix interactions, drug toxicology, and corneal regeneration.
    • The described methods facilitate advanced research in corneal physiology and disease using human-derived in vitro models.