Lung

LF/TERT309

Evercyte ́s human lung fibroblast cell line LF/TERT309 can be grown without limitations while maintaining expression of cell type specific markers and functions. Therefore, these cells are valuable as standardized in vitro model to study pulmonary function and pathophysiology such as airway inflammation and remodeling or production of extracellular matrix compounds.

General information

Cat#: CHT-067-0309

Organism: homo sapiens
Tissue, cell type: lung, 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

LF/TERT309 cells are characterized by the typical, spindle shaped morphology of fibroblastoid cells and homogenously express the fibroblast marker Vimentin. Cell nuclei are counterstained with DAPI.

Growth characteristics

LF/TERT309 cells can be grown for a minimum of 60 population doublings with a stable growth rate and without showing signs of growth retardation.

Myofibroblast differentiation / induction alpha SMA expression

Treatment of telomerized human lung fibroblasts LF/TERT309 with transforming growth factor beta (TGF-ß) induces expression of alpha smooth muscle actin (𝛂-SMA), indicating myofibroblast differentiation.

Treatment of telomerized human lung fibroblasts LF/TERT309 with transforming growth factor beta (TGF-ß) together with PP2, an inhibitor of Src familiy of tyrosine kinases, attenuates the response of the cells to TGF-ß.

In vitro propagation

DMEM/Ham´s F12 with FBS and G418

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

10 % FBS (PAN Biotech, Cat# P30-3031)

200 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 LF/TERT309
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.
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) and aspirate the cells by pipetting.
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 – 1:6 twice a week is recommended (after cells have reached about 70-90 % 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 % FBS (PAN Biotech, Cat# P30-3031)

10 % DMSO (Sigma Aldrich, Cat# D2650)

Additional material & reagents

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

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

Protocol cryopreservation of LF/TERT309
Freezing of cells

Detach the cells from the culture vessel by using Trypsin-EDTA solution (Protocol passaging of LF/TERT309)

Resuspend the detached cells in growth medium and centrifuge at 180 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 180 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 LF/TERT309

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 LF/TERT309 cell culture medium
Protocol passaging of LF/TERT309
Protocol cryopreservation of LF/TERT309
Immunofluorescence staining for vimentin expression

 

Data on Markers and Functions

LF/TERT309 – morphology and expression of vimentin
LF/TERT309 – myofibroblast differentiation upon treatment with TGFß

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 3 months: EUR 2000
Annual license fee R&D: royalty based
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LF/TERT309
Cat#: CHT-067-0309

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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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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,–

LF/TERT309
Cat#: CHT-067-0309

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO