Adipose tissue

ASC/TERT1

Evercyte ́s human adipose-derived mesenchymal stem cell line ASC/TERT1 can be grown without limitations while maintaining expression of cell type specific markers and functions. The cells are characterized by self-renewal and multipotent differentiation capacity. Therefore, the cell line is a relevant and standardizable in vitro model to study processes such as differentiation, inflammation as well as tissue homeostasis and repair.

General information

Cat#: CHT-001-0005-B

Organism: homo sapiens
Tissue, cell type: adipose tissue / liposuction (female donor), mesenchymal stem cells
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 growth characteristics

ASC/TERT1 cells are characterized by the typical spindle-shaped morphology of mesenchymal stem cells and can be grown for a minimum of 100 population doublings without showing signs of growth retardation. The cells show a constant growth rate with a population doubling time of 36-48 hours.

Expression of mesenchymal stem cell marker protein

ASC/TERT1 cells homogenously express typical marker proteins of mesenchymal stem cells such as CD73, CD90 and CD105, whereas the hematopoietic stem cell marker CD34 is not expressed (green peaks). Cells stained with isotype control antibodies are used as negative control (red peaks).

Differentiation potential towards adipocytes, osteoblasts and chondrocytes

ASC/TERT1 cells can be induced to differentiate towards adipocytes (oil red O staining), osteoblasts (Alizarin red staining) and chondrocytes ( alcian blue staining).

FAQs

In vitro propagation

Endothelial Cell Growth Medium-2  (Lonza, Cat# CC-3162) supplemented with FBS and G418

EBM-2 basal medium (Lonza, Cat# CC-3156)

Components of EGM-2 SingleQuot Kit (Lonza, Cat# CC-4176: Hydrocortisone, hFGF, VEGF, R3-IGF-1, Ascorbic acid, hEGF, Heparin)

2 % FBS (Sigma Aldrich, Cat# F7524)

200 µg/ml G418 (InvivoGen, Cat# ant-gn5)

Additional material & reagents

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

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

Defined Trypsin-Inhibitor (Gibco, Cat# R007100)

Protocol passaging of ASC/TERT1
Passaging of cells

For detachment of the cells remove and discard the culture medium and wash the cells twice with PBS (each 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 2-3 min.

Observe cell detachment under an inverted microscope. As soon as 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 1:3 – 1:4 twice a week is recommended (after cells have reached about 80 % confluence).
Cultivate cells at 37°C in a humidified atmosphere with 5% CO2.

Cryopreservation

Freezing medium

Complete growth medium (EGM-2 with supplements)

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

10 % DMSO (Sigma Aldrich, Cat# D2650)

Additional material & reagents

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

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

Defined Trypsin-Inhibitor (Gibco, Cat# R007100)

Protocol cryopreservation of ASC/TERT1
Freezing of cells

Detach the cells from the culture vessel by using trypsin-EDTA and trypsin-inhibitor (Protocol passaging of ASC/TERT1).

Resuspend the detached cells in complete 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

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 ASC/TERT1

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 ASC/TERT1 growth medium
Protocol passaging of ASC/TERT1
Protocol cryopreservation of ASC/TERT1

Data on Markers and Functions
ASC/TERT1 – morphology and growth characteristics
ASC/TERT1 – expression of typical MSC marker proteins CD73, CD90, CD105
ASC/TERT1 – potential to differentiate towards adipocytes, osteoblasts and chondrocytes
ASC/TERT1 – NGS data of undifferentiated cells generated by the Human Protein Atlas

Safety documents (coming soon)

Telomerized human cells – material safety data sheet
ASC/TERT1 – cell line establishment, vector maps

Selected publications

Fürsatz M, Gerges P, Wolbank S, Nürnberger S. Autonomous spheroid formation by culture plate compartmentation. Biofabrication. 2021 Jan 29. https://pubmed.ncbi.nlm.nih.gov/33513590.

Nürnberger S, Schneider C, Keibl C, Schädl B, Heimel P, Monforte X, Teuschl AH, Nalbach M, Thurner PJ, Grillari J, Redl H, Wolbank S. Repopulation of decellularised articular cartilage by laser-based matrix engraving. EBioMedicine. 2021 Feb;64:103196. https://pubmed.ncbi.nlm.nih.gov/33483297/
Katz DB, Huynh NPT, Savadipour A, Palte I, Guilak F. An immortalized human adipose-derived stem cell line with highly enhanced chondrogenic properties. Biochem Biophys Res Commun. 2020 Sep 10;530(1):252-258. https://pubmed.ncbi.nlm.nih.gov/32828295/
Comas F, Latorre J, Ortega F, Oliveras-Cañellas N, Lluch A, Ricart W, Fernández-Real JM, Moreno-Navarrete JM.
Permanent cystathionine-β-Synthase gene knockdown promotes inflammation and oxidative stress in immortalized human adipose-derived mesenchymal stem cells, enhancing their adipogenic capacity. Redox Biol. 2020 Aug 2:101668. https://pubmed.ncbi.nlm.nih.gov/32800520/
Wolbank S, Stadler G, Peterbauer A, Gillich A, Karbiener M, Streubel B, Wieser M, Katinger H, van Griensven M, Redl H, Gabriel C, Grillari J, Grillari-Voglauer R. Telomerase immortalized human amnion- and adipose-derived mesenchymal stem cells: maintenance of differentiation and immunomodulatory characteristics. Tissue Eng Part A. 2009 Jul;15(7):1843-54. https://pubmed.ncbi.nlm.nih.gov/19125642/
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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ASC/TERT1
Cat#: CHT-001-0005

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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.

“We have been using ASC/TERT1 since years for chondrogenic differentiation in in vitro as well as in vivo experiments. They are a valuable tool for avoiding donor variability allow for clearer and more constant results which is especially important for initial screenings or animal studies.”

 

Sylvia Nürnberger, principal investigator, Medical University of Vienna, Austria

Good cell culture practice / GCCP

get ready to standardize your cell culture work

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standardized production of spheroids for tissue engineering

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Cell factory for EV production

ASC/TERT300

human adipose-derived mesenchymal stem cells for production of clinical grade extracellular vesicles – coming soon

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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,–

ASC/TERT1
Cat#: CHT-001-0005

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO