Umbilical cord – vascular system

HUVEC/TERT2

Evercyte ́s human umbilical vein endothelial cell line HUVEC/TERT2 can be grown without limitations while maintaining expression of cell type specific markers and function. Therefore, these cells are useful for setting up in vitro bioassays for studying angiogenesis in e.g. wound healing or metastasis as well as bioassays for testing the angiogenic properties of novel drugs. Additionally, the cell line is the perfect starting material for genetic engineering to create important disease models.

General information

Cat#: CHT-006-0008

Organism: homo sapiens
Tissue, cell type: Umbilical cord, umbilical vein endothelial cells
Morphology:
endothelial 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 marker expression

HUVEC/TERT2 cells are characterized by the typical endothelial morphology and homogenous expression of cell-type specific markers such as vWF and CD31. Cell nuclei are counterstained with DAPI.

Response to VEGF treatment

Vascular endothelial growth factor (VEGF) induces proliferation in HUVEC/TERT2 cells in a concentration dependent manner (MTT assay).

Neo-angiogenic potential

When inoculated onto Matrigel-matrix HUVEC/TERT2 cells form typical capillary like structures (left picture) demonstrating neoangiogenic potential, which is also mirrored by the formation sprouts from 3D spheroids upon VEGF treatment (right picture).

Cellular proliferation upon drug treatment

HUVEC/TERT2 cells grown in multi-well plates are treated with vascular endothelial cell growth factor (VEGF) alone or in combination with anti-angiogenic drugs followed by analysis of cellular proliferation. A concentration dependent inhibition of growth promoting activity of VEGF by anti-angiogenic drugs lucentis® / Ranibizumab and avastin® / Bevacizumab is detected.

Sprout formation upon drug treatment

HUVEC/TERT2 cells are grown as spheroids in hanging drops followed by embedment into a semi-solid matrix. After treatment of these spheroids with vascular endothelial cell growth factor (VEGF), formation of sprouts is induced, which is inhibited by cultivation in the presence of cyclosporin A (CSA) in a concentration dependent manner.

FAQs

In vitro propagation

Endothelial growth medium (Lonza) supplemented with EGM SingleQuot Kit, FBS and G418

EBM basal medium (Lonza, Cat#CC-3121)

10 ng/ml hEGF (Sigma Aldrich, Cat# E9644)

Components of EGM SingleQuot Kit (Lonza, Cat# CC-4133: BBE, hEGF, hydrocortisones, ascorbic acid)

10 % FBS (Sigma Aldrich, Cat# F7524)

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

Additional material & reagents

0,1 % Gelatin (Sigma, Cat# G1393, 2 %), diluted in PBS

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

Trypsin inhibitor (Gibco, Cat# R007100)

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

Protocol passaging of HUVEC/TERT2
Passaging of cells

The new culture flasks have to be pre-coated with gelatin. Therefore, the culture flasks are treated with 0.1 % gelatin solution (60 µl/cm²) at 37°C for at least 10 min (10 – 60 min). Before introducing cells, remove excess of gelatin solution.

For detachment of the cells remove and discard the culture medium and wash the cells once with PBS (160 µl/cm²). Remove PBS completely.
Then, add 0.05 % Trypsin-EDTA (1x) 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 3 min.
Observe cell detachment under an inverted microscope. As soon as all cells are detached (if necessary, agitate the cells by gently hitting the flask), add growth medium (about 160 µl/cm²).

Add appropriate aliquots of the cell suspension to gelatin pre-coated culture vessels supplemented with growth medium (final volume of 240 µl/cm²).
A split ratio of 1:8 twice a week is recommended (after having reached about 90-95 % confluence).
Cultivate cells at 37°C in a humidified atmosphere with 5% CO2.

Cryopreservation

Freezing medium

Endothelial cell growth medium 10 % DMSO (Sigma Aldrich, Cat# D2650)

Additional material & reagents

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

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

0,1 % Gelatin (Sigma, Cat# G1393, 2 %), diluted in PBS

Protocol cryoprservation of HUVEC/TERT2
Freezing of cells

Detach the cells from the culture vessel by using Trypsin-EDTA solution (Protocol passaging of HUVEC/TERT2).

Resuspend the detached cells in endothelial cell 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 5 x 105 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 T75 rouxflask

Before thawing the original vial containing Evercyte cells, pre-coat a 25 cm² culture flask with gelatin (Protocol passaging of HUVEC/TERT2).
Add 6 ml of complete endothelial cell 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 near confluent at this point, they have to be passaged.
Protocol passaging of HUVEC/TERT2

Product data sheet – certificate of analysis Leaflet

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

Protocols

Preparation of endothelial cell growth medium
Protocol passaging of HUVEC/TERT2
Protocol cryopreservation of HUVEC/TERT2
Protocol for differentiation of HDMVEC/TERT164-B on Matrigel
Protocol immunofluorescent staining of HDMVEC/TERT164-B for marker expression

Data on Markers and Functions

HUVEC/TERT2 – morphology and marker expression
HUVEC/TERT2 – response to VEGF treatment
HUVEC/TERT2 – neoangiogenic potential
HUVEC/TERT2 – cellular proliferation upon drug treatment
HUVEC/TERT2 – sprout formation upon drug treatment
HUVEC/TERT2 – NGS data of undifferentiated cells generated by The Human Protein Atlas

Safety documents

Telomerized human cells – material safety data sheet
HUVEC/TERT2 – cell line establishment, vector maps

Selected publications

De Martin S, et al. (2021) Refill liquids for electronic cigarettes display peculiar toxicity on human endothelial cells. Toxicol Rep. 2021 Feb 26;8:456-462. https://pubmed.ncbi.nlm.nih.gov/33717998/

Gludovacz E, et al. (2020) Human diamine oxidase cellular binding and internalization in vitro and rapid clearance in vivo are not mediated by N-glycans but by heparan sulfate proteoglycan interactions. Glycobiology, cwaa090. https://pubmed.ncbi.nlm.nih.gov/32985651.

Noël JC, et al. (2020) Quantification of selected aroma compounds in e-cigarette products and toxicity evaluation in HUVEC/Tert2 cells. Biomed Chromatogr. 2020 Mar;34(3):e4761. https://pubmed.ncbi.nlm.nih.gov/31758585

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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Cat#: CHT-006-0008

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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.

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toxicity studies

HUVEC/TERT2 cells are useful to evaluate toxicity of aroma compounds in e-cigarette products

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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.

Print Friendly, PDF & Email

from

€ 1300,–

HUVEC/TERT2
Cat#: CHT-006-0008

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO