HCE-T

HCE-T is an SV40-transformed human corneal epithelial cell line derived from primary human corneal epithelium. The line was established by infection with a recombinant SV40–adenovirus hybrid vector (Ad–SV40), enabling stable expression of SV40 large T antigen and continuous proliferation. The original characterization specifically aimed to generate a continuously growing corneal epithelial cell line without shedding free viral particles.

In culture, HCE-T cells show typical epithelial “cobblestone” morphology and grow as adherent monolayers. Ultrastructural epithelial features such as desmosomes and apical microvilli have been reported, and the cells have been described as producing a cornea-associated 64 kD keratin. Under suitable differentiation conditions (e.g., air–liquid interface culture on collagen), HCE-T cells can form multilayered, stratified structures and develop measurable barrier properties, supporting their use in ocular surface research.

HCE-T cells are widely used to study corneal epithelial barrier function, permeability and formulation effects, migration/repair-related processes, and cellular responses to inflammatory or irritant stimuli. However, transporter expression patterns and differentiation-marker profiles can differ from native human cornea and from primary limbal/corneal epithelial systems. Therefore, HCE-T is best suited for mechanistic and comparative in vitro studies, while direct quantitative extrapolation to in vivo human corneal absorption or corneal differentiation biology should be performed with caution.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.