Cornea

hTCEpi

The cornea is constantly exposed to the outside environment and thus subject to damage due to various insults such as infections, drug- or chemical induced toxicity. In order to sustain such damage, the corneal epithelial cells form a resistance barrier, which is continuously renewed by stem cells located in the adjacent limbal region.

Evercyte offers a continuously growing corneal epithelial cell line with functions that resemble that of the corresponding primary counterpart cells. Therefore, our cells qualify for studying drug transport, drug- and chemical-induced toxicity on the corneal epithelium as well as inflammation processes and wound healing.

General information

Cat#: CHT-045-0237

Organism: homo sapiens
Tissue, cell type: limbal region of the corneal tissue, corneal epithelial cells
Morphology:
epithelial morphology
Life span extension:
ectopic expression of hTERT
Quality:
free from contaminations (bacteria incl. mycoplasma, fungi, HIV, HAV, HBV, HCV, Parvo-B19) and cross-contaminations

Morphology and expression of marker proteins

hTCEpi cells are characterized by the typical epithelial cobblestone appearance and expression of corneal epithelial cell markers such as ZO1 and KRT3.

FAQs

In vitro propagation

Keratinocyte growth medium KGM-2 Bullet Kit (Lonza)

KBM-2 basal medium (Lonza, Cat# CC-3103)

Components of KGM-2 SingleQuot Kit (Lonza, Cat# CC-4152: BPE, hEGF, insulin, hydrocortisone, transferrin, epinephrine)

Additional material & reagents

Phosphate buffered saline (PBS) (Sigma, Cat# D8537)

Trypsin inhibitor (Gibco, Cat# R007100)

0,05 % Trypsin-EDTA (Gibco, Cat#25300-054)

Protocol passaging of hTCEpi
Passaging of cells

For detachment of the cells remove and discard the culture medium and wash the cells once with PBS (about 160 µl/cm²). Remove PBS completely.

Then, add 0.05 % Trypsin-EDTA solution (20 µl/cm²), make sure that all cells have been in contact with this solution and incubate the culture flask at 37°C for approximately 4-5 min.
Observe cell detachment under an inverted microscope. Cells typically detach as a cell layer which decomposes when resuspended in medium.
As soon as the cells are detached, add Trypsin-Inhibitor (20 µl/cm²), resuspend the cells in growth medium (about 160 µl/cm²) and centrifuge at 170 g for 5 min.
Discard the supernatant, resuspend the cell pellet in the remaining droplet and add growth medium (about 160 µl/cm²).
Then, transfer appropriate aliquots of the cell suspension to new culture vessels supplemented with growth medium (final volume of 240 µl/cm²).
A split ratio of 1:8 twice a week is recommended (after cells have reached about 60-70 % confluence).
Never allow the culture to become confluent!
Cultivate cells at 37°C in a humidified atmosphere with 5% CO2.

Cryopreservation

Freezing medium

CryoStor® cell cryopreservation medium CS10 (Sigma Aldrich, Cat# C2874)

Additional material & reagents

Phosphate buffered saline (PBS) (Sigma, Cat# D8537)

0,05 % Trypsin-EDTA (Gibco, Cat# 25300-054)

Trypsin inhibitor (Gibco, Cat# R007100)

Protocol cryopreservation of hTCEpi
Freezing of cells

Detach the cells from the culture vessel by using Trypsin-EDTA and Trypsin-Inhibitor (Protocol passaging of hTCEpi).

Resuspend the detached cells in KGM-2 growth medium and centrifuge at 170 g for 5 min.
Discard the supernatant, resuspend the resulting cell pellet in the remaining droplet and add freezing medium (tempered to 4°C) to reach a cell density of about 1 x 106 cells/ml (for thawing in a 25 cm² culture flask).
Add 1 ml of this cell suspension to each pre-cooled cryovial and immediately transfer the cells to -80°C.
After 24 hours transfer the vials to the liquid nitrogen tank.
Thawing of cells

Original Evercyte cells are to be thawed in a T5 rouxflask

Add 6 ml of complete KGM-2 growth medium to a 25 cm² culture flask and place the culture flask in the incubator for at least 30 min to allow the medium to reach its normal pH.
Take a vial of frozen cells, rinse it outside with Ethanol and pre-warm in the hand until one last piece of frozen cells is seen.
Then, immediately transfer the content of the vial to a 15 ml centrifugation tube pre-filled with 9 ml of medium pre-cooled to 4°C and centrifuge for 5 min at 170 g.
Discard the supernatant and resuspend the cell pellet in the remaining droplet.
Add 1 ml of the pre-warmed medium to the cells, transfer them to the prepared culture flask and incubate at 37°C in a suitable incubator.
Perform a medium change 24 hours after thawing. If the cells are already about 60-70 % confluent at this point, they have to be passaged.
Protocol passaging of hTCEpi

Product data sheet – certificate of analysis

Product data sheet (PDS) download
Certificate of analysis
is available upon request | Please contact us indicating the respective LOT numbers

Protocols

Preparation of hTCEpi cell culture medium
Protocol passaging of hTCEpi
Protocol cryopreservation of hTCEpi

Data on Markers and Functions

hTCEpi – morphology and marker expression

Safety documents (coming soon)

Telomerized human cells – material safety data sheet

hTCEpi – cell line establishment, vector maps

Selected publications

Raghunathan V, Edwards SG, Leonard BC, Kim S, Evashenk AT, Song Y, Rewinski E, Marangakis Price A, Hoehn A, Chang C, Reilly CM, Muppala S, Murphy CJ, Thomasy SM.  Differential effects of Hsp90 inhibition on corneal cells in vitro and in vivo. Exp Eye Res. 2021 Jan;202:108362.  https://pubmed.ncbi.nlm.nih.gov/33220237/

Kasus-Jacobi A, Land CA, Stock AJ, Washburn JL, Pereira HA.  Antimicrobial Peptides Derived from the Immune Defense Protein CAP37 Inhibit TLR4 Activation by S100A9.  Invest Ophthalmol Vis Sci. 2020 Apr 9;61(4):16.  https://pubmed.ncbi.nlm.nih.gov/32298435/

Alekseev O, Donegan WE, Donovan KR, Limonnik V, Azizkhan-Clifford J. HSV-1 Hijacks the Host DNA Damage Response in Corneal Epithelial Cells through ICP4-Mediated Activation of ATM.  Invest Ophthalmol Vis Sci. 2020 Jun 3;61(6):39.  https://pubmed.ncbi.nlm.nih.gov/32543665/

Kim S, Gates B, Leonard BC, Gragg M, Pinkerton KE, Winkle LV, Murphy CJ, Pyrgiotakis G, Zhang Z, Demokritou P, Thomasy SM. Engineered metal oxide nanomaterials inhibit corneal epithelial wound healing in vitro and in vivo. NanoImpact. 2020 Jan;17:100198.  https://pubmed.ncbi.nlm.nih.gov/32154443/

Robertson DM, Li L, Fisher S, Pearce VP, Shay JW, Wright WE, Cavanagh HD, Jester JV. Characterization of growth and differentiation in a telomerase-immortalized human corneal epithelial cell line. Invest Ophthalmol Vis Sci. 2005 Feb;46(2):470-8. https://pubmed.ncbi.nlm.nih.gov/15671271/

List of publications

Licence Conditions

The business concept of Evercyte is to out-license telomerized cells to our customers. The license conditions depend on whether the contract partner is a for profit or a nonprofit organization and the intended use of the cells.

Nonprofit organizations

Evercyte grants licenses for an unlimited period to academic or nonprofit-organizations, whereby the use of Evercyte cell lines is restricted to research & development purposes and non-commercial use. The cells are not intended for human use.
The customers have to agree to the conditions described in our material transfer agreement as well as accept our general terms and conditions.
On time payment for unlimited use: EUR 1300

Profit organizations

Pharmaceutical – chemical- cosmetic industries
Evercyte grants licenses for commercial organizations, whereby we offer an initial testing phase for a flat fee that allows our customers to test our cells in their laboratories for a period of 6 months.
Thereafter, annual license fees fall due, depending on the cell line of interest. Besides offering cell lines for research & development purposes, we also have established cell factories that qualify for production of clinical grade extracellular vesicles for human application.
The customer has to agree to the conditions described in our license agreements.
Contract research organizations (CRO)
Evercyte grants licenses for contract research organizations, whereby we offer an initial testing phase for a flat fee that allows our customers to test our cells in their laboratories. Thereafter, we would negotiate a royalty based long-term license agreement individually.
The use of the cells during these phases is restricted to research & development purposes. The cells are not intended for human use. The customers have to agree to the conditions described in our material transfer agreement and accept our general terms and conditions.
Initial license fee for 6 months: EUR 2000
Annual license fee R&D: royalty based
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Customer Reviews

“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”

 

Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.

“Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform.  We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed.”

Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.

about
our cells

telomerized (hTERT immortalized) cells in biotechnology and biomedicine

read more

study of inflammation

hTCEpi cells are fit for purpose to study inflammation in dry eye disease

read more

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Related Services

specific request

Customer Reviews

“I have had the pleasure of working with Evercyte for the last few years. We continually rely on Evercyte because of the high-quality data that they produce, their diligent responsiveness, and their excellent customer service.”

 

Josh Garlich, Senior Research Scientist, Apellis Pharmaceuticals, Inc.

“Cytonus has been working with Evercyte from many years as they are a trusted partner and have always delivered the highest quality cell lines to advance our platform.  We routinely draw on their expertise to meet cellular engineering challenges and they have not disappointed.”

Remo Moomiaie-Qajar, Cytonus Therapeutics, Inc.

Print Friendly, PDF & Email

from

€ 1300,–

hTCEpi
Cat#: CHT-045-0237

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO