fHDF/TERT1662021-08-02T06:43:08+00:00

Skin

fHDF/TERT166

Evercyte ́s human dermal fibroblast cell line fHDF/TERT166 can be grown without limitations while maintaining expression of cell type specific markers and functions. Therefore, these cells are frequently used as standardized in vitro model to study processes involving fibroblasts such as formation of the extracellular matrix, inflammation, wound healing or fibrosis. Moreover, the cells embedded into a collagen matrix and co-cultured with telomerized keratinocytes allow the establishment of standardizable 3D skin equivalents.

General information

Cat#: CHT-031-0166

Organism: homo sapiens
Tissue, cell type: foreskin (male), fibroblasts
Morphology:
mesenchymal 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

fHDF/TERT166 cells are characterized by the typical, spindle shaped morphology of mesenchymal cells and homogenously express the fibroblast marker Vimentin. Cell nuclei are counterstained with DAPI.

Response to cytokines / expression of IL-6 upon IL-17A treatment

Treatment of telomerized human dermal fibroblasts fHDF/TERT166 with Interleukin-17A (IL17A) results in expression of interleukin-6 (IL6) in a concentration dependent manner.

Myofibroblast differentiation / effect of extracellular vesicles

Treatment of telomerized human dermal fibroblasts fHDF/TERT166 with transforming growth factor beta (TGF-ß) induces expression of alpha smooth muscle actin (𝛂-SMA). Treatment of cells with TGF-ß together with extracellular vesicles (EVs) from Wharton´s Jelly derived mesenchymal stem cells (WJ-MSC/TERT273) reduces 𝛂-SMA expression significantly, demonstrating an effect of EVs on myofibroblast differentiation.

Cell migration / induction of fibroblast growth upon treatment with extracellular vesicles

fHDF/TERT166 were seeded into chamber slides and a physical gab within the monolayer was created followed by monitored the process of cell migration into the gap. Significantly less free area between the cells was detected upon addition of extracellular vesicles from MSCs (WJ-MSC/TERT273).

FAQs

What is the difference between fHDF/TERT166 and HDF/TERT164 cells?2021-06-21T07:54:24+00:00

fHDF/TER166 cells are derived from foreskin, whereas HDF/TERT164 cells are from adult skin (Cutis Laxa). Both cell lines show typical markers and characteristics of fibroblastoid cells and a stable growth rate. However, fHDF/TERT166 cells show a higher growth rate (population doubling time of 48 hours) compared to HDF/TERT164 cells (population doubling time of 60-72 hours).

In vitro propagation

DMEM/Ham´s F12 supplemented with Fetal Bovine Serum and G418

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DMEM/Ham´s F12 (1:1) (PAN Biotech, Cat# P04-41150)

10 % FBS (Sigma-Aldrich, Cat# F7524)

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

Additional material & reagents

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

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

Protocol passaging of fHDF/TERT166
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 3 min.
Add appropriate aliquots of the cell suspension to new culture vessels supplemented with growth medium (final volume of 240 µl/cm²).

A split ratio of 1:4 twice a week is recommended (after cells have reached about 90 % confluence).

Cultivate cells at 37°C in a humidified atmosphere with 5% CO2.

Cryopreservation

Freezing medium

DMEM/Ham´s F12 (1:1) (PAN Biotech, Cat# P04-41150)

10 % Fetal bovine serum (Sigma Aldrich, Cat# F7524)

10 % DMSO (Sigma Aldrich, Cat# D2650)

Additional material & reagents

Phosphate buffered saline (PBS) (Sigma, Cat# D8537)[ fusion_text]

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

Protocol cryopreservation of fHDF/TERT166
Freezing of cells

Detach the cells from the culture vessel by using Trypsin-EDTA solution (Protocol passaging of fHDF/TERT166)

Resuspend the detached cells in 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 10^5 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 T25 roux flask

Add 6 ml of complete 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 80 % confluent at this point, they have to be passaged.
Protocol passaging of fHDF/TERT166

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 fHDF/TERT166 cell culture medium
Protocol passaging of fHDF/TERT166
Protocol cryopreservation of fHDF/TERT166

Data on Markers and Functions

fHDF/TERT166 – morphology and expression of vimentin
fHDF/TERT166 – response to Interleukin-17A treatm.
fHDF/TERT166 – myofibroblast differentiation upon treatment with extracellular vesicles
fHDF/TERT166 – scratch assay, cell migration upon treatment with extracellular vesicles

Safety documents (coming soon)

Telomerized human cells – material safety data sheet
fHDF/TERT166 – cell line establishment, vector maps

 

Selected publications

Coming soon

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-031-0166

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“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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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,–

fHDF/TERT166
Cat#: CHT-031-0166

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO

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