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Step-by-Step Guide to Corneal Epithelial Cell Culture Products

Human Corneal Epithelial Cells

Our Gibco® Human Corneal Epithelial Cells (HCECs) are normal corneal epithelial cells isolated from the progenitor-rich limbal region of the eye. Primary HCECs are prepared to provide ≥70% viability upon thawing, with each vial containing sufficient cells to seed =100 cm2 of tissue culture surface.

In this picture, HCECs are imaged using the Click-iT® Edu Alexa Fluor® 488 imaging kit, anti- alpha -tubulin antibody with Goat anti-mouse Alexa Fluor® 555 secondary, and Hoechst 33342.

Each lot of HCECs undergoes performance testing and is guaranteed to achieve at least 12 population doublings (PD) after thawing when using Keratinocyte Serum-Free Medium (KSFM). HCECs stain positive in immunocytochemistry screens for the corneal epithelial markers cytokeratin 15 and p63 alpha.

Figure 1. Phase contrast image of HCEC at days 0, 2, and 4. Typically, HCEC reach confluency at day 4-6 when cultured according to product instructions in Keratinocyte-SFM.

 

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Corneal Media Systems

Optimized for use with Gibco® media.
Invitrogen™ Human Corneal Epithelial Cells (HCECs) have been optimized for use with our serum-free media, specifically, Gibco® Keratinocyte Serum-Free Medium (KSFM) or Gibco® Defined Keratinocyte Serum-Free Medium (DKSFM), when used in combination with Coating Matrix Kit for applications where a defined growth system is important. Preparation of supplemented media is a simple process, and the final solution can be stored for up to 30 days at 4°C protected from light.

 

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Figure 2. HCECs were thawed and seeded according to product instructions provided with Keratinocyte SFM (KSFM) or Defined Keratinocyte SFM (DKSFM). Cells were passaged at ~90% confluence, and population doublings per culture were calculated. Bars show the mean population doubling of triplicate T-25 flasks with standard deviation.

 

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Media and Reagents

Alternative Media and Supplements

Recommended Reagents

For optimal performance when culturing human corneal epithelial cells, we recommend using the cell dissociation, coating, and cryopreservation reagents found on this page.

In this picture, HCECs are labeled with the Alexa Fluor® 488 phalloidin and an anti–a-tubulin antibody followed by Alexa Fluor® 555 goat anti–mouse IgG secondary antibody. HCECs were counterstained with HCS CellMask™ Blue Stain.

Our human corneal epithelial cells also work well with a range of Invitrogen™ and Molecular Probes® cellular imaging and analysis tools.

 

Figure 3. HCEC imaged using BacMam CellLight® ER-GFP and CellLight® Talin-RFP,  and Hoechst 33342 (left image). HCEC imaged using BacMam CellLight® ER-GFP  and Hoechst 33342 (right image).

 

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Cell Dissociation

Coating

Cryopreservation

Imaging

 

Undefined Growth System Defined Growth System

 

Resources

Life Technologies provides a comprehensive set of resources for primary cell research, including protocols and technical support.

Use the links (right) to browse our current information on corneal cell culture.

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