Skin

NHEK/SVTERT3-5

NHEK/SVTERT3-5 cells show an extended in vitro life span and express typical functions of skin keratinocytes as they can be differentiated towards 3D skin equivalents in an air-liquid interface. Thus, the cell line can be used as standardized in vitro model to study processes such as wound healing or to investigate the uptake of certain drugs. Additionally, due to its unique characteristics, the cell line allows studying chronic intoxication as demonstrated using arsenic.

General information

Cat#: CLHT-011-0026-5

Organism: homo sapiens
Tissue, cell type: skin, epidermal keratinocytes
Morphology:
epithelial morphology
Life span extension:
ectopic expression of hTERT and SV40 largeT antigen
Quality:
free from contaminations (bacteria incl. mycoplasma, fungi, HIV, HAV, HBV, HCV, Parvo-B19) and cross-contaminations

Morphology and 3D differentiation

NHEK/SVTERT3-5 cells show the typical cobblestone epithelial morphology of keratinocytes and can be differentiated towards a stratified skin equivalent in an air-liquid interface.

FAQs

In vitro propagation

[/fusion_text]

Keratinocyte growth medium KGM-2 Bullet Kit (Lonza, Cat# CC-3107)

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

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

20 µg/ml G418 (InvivoGen, Cat# ant-gn-5)

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 NHEK/SVTERT3-5
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. 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 1:2 – 1:4 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 cryoprservation of NHEK/SVTERT3-5
Freezing of cells

Detach the cells (60-70 % confluence) from the culture vessel by using Trypsin-EDTA and Trypsin-Inhibitor (Protocol passaging of NHEK/SVTERT3-5).

Resuspend the detached cells in growth medium and centrifuge at 170 g for 5 min.
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 T25 rouxflask

Add 6 ml of 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.
Recovery from cryopreservation will take a few days. 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 NHEK/SVTERT3-5

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 NHEK/SVTERT3-5 cell culture medium
Protocol passaging of NHEK/SVTERT3-5
Protocol cryopreservation of NHEK/SVTERT3-5

Data on Markers and Functions

NHEK/SVTERT3-5 – morphology and 3D differentiation

Safety documents (coming soon)

Telomerized human cells – material safety data sheet

NHEK/SVTERT3-5 – cell line establishment, vector maps

Publications (telomerized keratinocytes from Evercyte)

Weinmuellner R, Kryeziu K, Zbiral B, Tav K, Schoenhacker-Alte B, Groza D, Wimmer L, Schosserer M, Nagelreiter F, Rösinger S, Mildner M, Tschachler E, Grusch M, Grillari J, Heffeter P. Long-term exposure of immortalized keratinocytes to arsenic induces EMT, impairs differentiation in organotypic skin models and mimics aspects of human skin derangements. Arch Toxicol. 2018 Jan;92(1):181-194. https://pubmed.ncbi.nlm.nih.gov/ 28776197 /

Wagner T, Gschwandtner M, Strajeriu A, Elbe-Bürger A, Grillari J, Grillari-Voglauer R, Greiner G, Golabi B, Tschachler E, Mildner M. Establishment of keratinocyte cell lines from human hair follicles. Sci Rep. 2018 Sep 7;8(1):13434. https://pubmed.ncbi.nlm.nih.gov/30194332/
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
Print Friendly, PDF & Email

from

€ 1300,–

NHEK-SVTERT3-5
Cat#: CLHT-011-0026-5

Ordering now

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO

Related Products

human skin fibroblasts fHDF/TERT166

read more

Related Services

send us 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.

study of long-term exposure to toxicants

arsenic impairs differentiation of NHEK/SVTERT3-5 cells in a 3D skin equivalent

read more

news

Lecture on 3Rs

(replacement – reduction -refinement of animal experiments)

read more

Related Products

MyoUp MHT-040

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,–

NHEK-SVTERT3-5
Cat#: CLHT-011-0026-5

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO