RPTEC/TERT12021-08-16T09:39:03+00:00

Kidney

RPTEC/TERT1

Evercyte ́s human renal proximal tubular epithelial cell line RPTEC/TERT1 can be grown without limitations while maintaining expression of cell type specific markers and function. Therefore, these cells are useful to study kidney transport functions, kidney diseases or diabetes and to test new drugs for cytotoxic effects. Additionally, the cell line is the perfect starting material for genetic engineering to create important disease models.

General information

Cat#: CHT-003-0002

Organism: homo sapiens
Tissue, cell type: kidney cortex, male donor, renal proximal tubular epithelial cells
Morphology:
epithelial, cobblestone 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

RPTEC/TERT1 cells are characterized by the typical epithelial morphology and formation of domes when grown to high cell densities, which demonstrates active transport of water from the apical to the basolateral side. Moreover, the cells form continuous belts of E-cadherin and ZO-1 at cell-cell contacts.

FAQs

Can you provide a protocol for transfection of the cells?2021-06-21T09:45:25+00:00

The cells can be transfected using Lipofectamine 2000 as described in Wieser M, et al. (2019) CD46 knock-out using CRISPR/Cas9 editing of hTERT immortalized human cells modulates complement activation. PLoS One. 2019; 14(4): e0214514. www.ncbi.nlm.nih.gov/pmc/articles/PMC6453361

Are RNA seq data available for RPTEC/TERT1 cell line?2021-06-21T09:43:49+00:00

NGS data of undifferentiated cells have been generated by the Human Protein Atlas ® (https://www.proteinatlas.org/search/RPTEC%2FTERT1)

In vitro propagation

DMEM/F12 (PAN Biotech, Cat # P04-41154)

10 mM HEPES-buffer (Sigma Aldrich, Cat# H0887)

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

5 pM 3,3′,5-Triiodo-L-thyronine sodium salt (Sigma Aldrich, Cat# T6397)

3.5 µg/ml L-Ascorbic Acid (Sigma Aldrich, Cat# A4544)

5 µg/ml Transferrin Holo (Merck Millipore, Cat# 616424)

25 ng/ml Prostaglandine E1 (Sigma Aldrich, Cat# P8908)

25 ng/ml Hydrocortisone (Sigma Aldrich, Cat# H0396)

8.65 ng/ml Sodium-Selenite (Sigma Aldrich, Cat# S5261)

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

5 µg/ml Insulin (Sigma Aldrich, Cat# I9278)

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 RPTEC/TERT1
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 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 – 5 min. Observe the cell detachment under an inverted microscope.
As soon as all cells are detached (if necessary, agitate the cells by gently hitting the flask), 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, 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 1:2 to 1:3 twice a week is recommended (after having reached about 95 % confluence). Cells should be passaged when near 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 cryopreservation of RPTEC/TERT1
Freezing of cells

Detach the cells from the culture vessel by using Trypsin and Trypsin-Inhibitor (Protocol passaging of RPTEC/TERT1).

Resuspend the detached cells in growth medium and centrifuge at 170 g for 5 min.
Discard the supernatant, resuspend in the remaining droplet and add freezing medium (4°C) to reach a cell density of about 1,5 – 2 x 10^6 cells/ml (for thawing in a 25 cm² culture flask).
Transfer 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 growth medium to a 25 cm² culture flask and place the culture flask in the incubator for at least 30 min.
Take a vial of frozen cells, rinse outside with Ethanol and pre-warm in 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 cells in the remaining droplet.
Add 1 ml of pre-warmed medium to the cells, transfer the cell suspension 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 confluent at this point, they have to be passaged. Typically, RPTEC/TERT1 cells can be split 2-3 days after thawing when having reached about 95% confluence.
Protocol passaging of RPTEC/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 cell culture medium ProxUp
Protocol passaging of RPTEC/TERT1
Protocol cryopreservation of RPTEC/TERT1
Protocol for measurement of gamma-glutamyl-transferase (GGT) activity
Protocol for staining of RPTEC/TERT1 cells for marker expression / E-Cadherin
Protocol for staining of RPTEC/TERT1 cells for marker expression / CD13
Protocol for staining of RPTEC/TERT1 cells for marker expression / ZO-1

Data on Markers and Functions

RPTEC/TERT1 – morphology, expression of E-cadherin and ZO-1
RPTEC/TERT1 – NGS data of undifferentiated cells generated by the Human Protein Atlas

Safety documents (coming soon)

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

Selected publications

Glykofridis IE, et al. (2021) Loss of FLCN-FNIP1/2 induces a non-canonical interferon response in human renal tubular epithelial cells. eLife. 2021; 10: e61630 www.ncbi.nlm.nih.gov/pmc/articles/PMC7899648/

Delp J, et al. (2021) Neurotoxicity and underlying cellular changes of 21 mitochondrial respiratory chain inhibitors. Arch Toxicol. 2021; 95(2): 591–615. www.ncbi.nlm.nih.gov/pmc/articles/PMC7870626/
Megan L, et al. (2021) Effects of Proximal Tubule Shortening on Protein Excretion in a Lowe Syndrome Model. J Am Soc Nephrol. 2020 Jan; 31(1): 67–83. www.ncbi.nlm.nih.gov/pmc/articles/PMC6934997/
Krebs A, et al. (2020) The EU-ToxRisk method documentation, data processing and chemical testing pipeline for the regulatory use of new approach methods. Arch Toxicol. 2020; 94(7): 2435–2461. www.ncbi.nlm.nih.gov/pmc/articles/PMC7367925/
Spinu N, et al. (2020) Quantitative adverse outcome pathway (qAOP) models for toxicity prediction. Arch Toxicol. 2020; 94(5): 1497–1510. www.ncbi.nlm.nih.gov/pmc/articles/PMC7261727/
van der Stel W, et al. (2020) Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals. Arch Toxicol. 2020; 94(8): 2707–2729. www.ncbi.nlm.nih.gov/pmc/articles/PMC7395062/
Shrestha S, et al. (2020) Characterization and determination of cadmium resistance of CD133+/CD24+ and CD133−/CD24+ cells isolated from the immortalized human proximal tubule cell line, RPTEC/TERT1. www.ncbi.nlm.nih.gov/pmc/articles/PMC6766375/
Obaidi I, et al. (2020) Curcumin Sensitizes Kidney Cancer Cells to TRAIL-Induced Apoptosis via ROS Mediated Activation of JNK-CHOP Pathway and Upregulation of DR4. Biology (Basel) 2020 May; 9(5): 92. www.ncbi.nlm.nih.gov/pmc/articles/PMC7284747/
Pirklbauer M, et al. (2020) Empagliflozin Inhibits Basal and IL-1β-Mediated MCP-1/CCL2 and Endothelin-1 Expression in Human Proximal Tubular Cells. Int J Mol Sci. 2020 Nov; 21(21): 8189. www.ncbi.nlm.nih.gov/pmc/articles/PMC7663377/
Wieser M, et al. (2019) CD46 knock-out using CRISPR/Cas9 editing of hTERT immortalized human cells modulates complement activation. PLoS One. 2019; 14(4): e0214514. www.ncbi.nlm.nih.gov/pmc/articles/PMC6453361/
Limonciel A, et al. (2018) Comparison of base-line and chemical-induced transcriptomic responses in HepaRG and RPTEC/TERT1 cells using TempO-Seq. Arch Toxicol. 2018; 92(8): 2517–2531. www.ncbi.nlm.nih.gov/pmc/articles/PMC6063331/
Kramer N.I., et al. (2015) Biokinetics in repeated-dosing in vitro drug toxicity studies. Toxicol In Vitro. 2015 Sep8. pii: S0887-2333(15)00218-0. https://pubmed.ncbi.nlm.nih.gov/26362508/
Slyne J, et al., (2105) New developments concerning the proximal tubule in diabetic nephropathy: in vitro models and mechanisms. Nephrol Dial Transplant. 2015 Aug;30. https://pubmed.ncbi.nlm.nih.gov/26209740/

Ranninger C., et al. (2015) Nephron Toxicity Profiling via Untargeted Metabolome Analysis Employing a High
Performance Liquid Chromatography-Mass Spectrometry-based Experimental and Computational Pipeline. J BiolChem. 2015 Jul 31;290(31):19121-32. https://pubmed.ncbi.nlm.nih.gov/26055719/

Simon-Friedt BR, et al. (2015) The RPTEC/TERT1 Cell Line as an Improved Tool for In Vitro Nephrotoxicity
Assessments. Biol Trace Elem Res. 2015 Jul;166(1):66-71. https://pubmed.ncbi.nlm.nih.gov/25893367/

Maschmeyer I, et al. (2015) A four-organ-chip for inter-connected long-term co-culture of human intestine, liver, skin and kidney equivalents. Lab Chip. 2015 Jun 21;15(12):2688-99.https://pubmed.ncbi.nlm.nih.gov/25996126/

Fliedl L, et al. (2015) Optimisation of a quantitative PCR based method for Plasmid Copy Number Determination in Human Cell Lines. N Biotechnol. 2015 Mar 18. pii: S1871-6784(15)00046-1. https://pubmed.ncbi.nlm.nih.gov/25796475/
Limonciel A., et al. (2015) Transcriptomics hit the target: Monitoring of ligand-activated and stress response
pathways for chemical testing. Toxicol In Vitro. 2015 Jan 13. pii: S0887-2333(14)00251-3. https://pubmed.ncbi.nlm.nih.gov/25596134/
Crean D., et al. (2014) Development of an in vitro renal epithelial disease state model for xenobiotic toxicity testing. Toxicol In Vitro. 2014 Dec 20. pii: S0887-2333(14)00239-2. https://pubmed.ncbi.nlm.nih.gov/25536518/

Aschauer L, et al. (2015) Application of RPTEC/TERT1 cells for investigation of repeat dose nephrotoxicity: A
transcriptomic study. Toxicol In Vitro. 2015 Dec 25;30(1 Pt A):106-16. https://pubmed.ncbi.nlm.nih.gov/25450743/
Wilmes A, et al. (2015) Mechanism of cisplatin proximal tubule toxicityrevealed by integrating transcriptomics, proteomics, metabolomics and biokinetics. Toxicol In Vitro. 2015 Dec 25;30(1 Pt A):117-27. https://pubmed.ncbi.nlm.nih.gov/25450742/

Wilmes A, et al. (2014) Evidence for a role of claudin 2 as a proximal tubular stress responsive paracellular water channel. Toxicol Appl Pharmacol. 2014 Sep 1;279(2):163-72. https://pubmed.ncbi.nlm.nih.gov/24907557/

Simon BR, et al. (2014) Cadmium alters the formation of benzo[a]pyrene DNA adducts in the RPTEC/TERT1 human renal proximal tubule epithelial cell line. Toxicol Rep. 2014 Jul 14;1:391-400. https://pubmed.ncbi.nlm.nih.gov/25170436/

Fliedl L, Wieser M, Manhart G, Gerstl MP, Khan A, Grillari J, Grillari-Voglauer R. (2014) Controversial role of gammaglutamyl transferase activity in cisplatin nephrotoxicity. ALTEX. 2014;31(3):269-78. doi: Epub 2014 Mar 21. https://pubmed.ncbi.nlm.nih.gov/24664430/

Aschauer L, et al. (2013) Delineation of the Key Aspects in the Regulation of Epithelial Monolayer Formation. Mol Cell Biol. 2013 Jul;33(13):2535-50. https://pubmed.ncbi.nlm.nih.gov/23608536/

Radford R, Slattery C, Jennings P, Blacque O, Pfaller W, Gmuender H, Van Delft J, Ryan MP, McMorrow T. (2012) Carcinogens induce loss of the primary cilium in human renal proximal tubular epithelial cells independently of effects on the cell cycle. Am J Physiol Renal Physiol. 2012 Apr 15;302(8):F905-16. doi: 10.1152/ajprenal.00427.2011. Epub 2012 Jan 18. https://pubmed.ncbi.nlm.nih.gov/22262483/

Wilmes A, Limonciel A, Aschauer L, Moenks K, Bielow C, Leonard MO, Hamon J, Carpi D, Ruzek S, Handler A, Schmal O, Herrgen K, Bellwon P, Burek C, Truisi GL, Hewitt P, Di Consiglio E, Testai E, Blaauboer BJ, Guillou C, Huber CG, Lukas A, Pfaller W, Mueller SO, Bois FY, Dekant W, Jennings P. (2013) Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress. J Proteomics. 2013 Feb 21;79:180-94. https://pubmed.ncbi.nlm.nih.gov/23238060/

Limonciel A, Wilmes A, Aschauer L, Radford R, Bloch KM, McMorrow T, Pfaller W, van Delft JH, Slattery C, Ryan MP, Lock EA, Jennings P. (2012) Oxidative stress induced by potassium bromate exposure results in altered tight junction protein expression in renal proximal tubule cells. Arch Toxicol. 2012 Nov;86(11):1741-51. doi: 10.1007/s00204-012-0897-0. Epub 2012 Jul 4. https://pubmed.ncbi.nlm.nih.gov/22760423/

Jennings P, Weiland C, Limonciel A, Bloch KM, Radford R, Aschauer L, McMorrow T, Wilmes A, Pfaller W, Ahr HJ, Slattery C, Lock EA, Ryan MP, Ellinger-Ziegelbauer H. (2012) Transcriptomic alterations induced by Ochratoxin A in rat and human renal proximal tubular in vitro models and comparison to a rat in vivo model. Arch Toxicol. 2012 Apr;86(4):571-89. doi: 10.1007/s00204-011-0780-4. Epub 2011 Nov 29. https://pubmed.ncbi.nlm.nih.gov/22124623/

Sarkozi R. et al. (2011) Oncostatin M is a novel inhibitor of TGF-β1-induced matricellular protein expression. Am J Physiol Renal Physiol 2011 Nov, 301(5):F1014-F1025. https://pubmed.ncbi.nlm.nih.gov/21816755/

Limonciel A, Aschauer L, Wilmes A, Prajczer S, Leonard MO, Pfaller W, Jennings P. (2011) Lactate is an ideal noninvasive marker for evaluating temporal alterations in cell stress and toxicity in repeat dose testing regimes. Toxicol In Vitro. 2011 Dec;25(8):1855-62. doi: 10.1016/j.tiv.2011.05.018. Epub 2011 May 24. https://pubmed.ncbi.nlm.nih.gov/21635945/

Ellis JK, Athersuch TJ, Cavill R, Radford R, Slattery C, Jennings P, McMorrow T, Ryan MP, Ebbels TM, Keun HC. (2011) Metabolic response to low-level toxicant exposure in a novel renal tubule epithelial cell system. Mol Biosyst. 2011 Jan;7(1):247-57. doi: 10.1039/c0mb00146e. Epub 2010 Nov 19. https://pubmed.ncbi.nlm.nih.gov/21103459/

Wieser M, Stadler G, Jennings P, Streubel B, Pfaller W, Ambros P, Riedl C, Katinger H, Grillari J, Grillari-Voglauer R. (2008) hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics. Am J Physiol Renal Physiol. 2008 Nov;295(5):F1365-75. doi: 10.1152/ajprenal.90405.2008. Epub 2008 Aug 20. https://pubmed.ncbi.nlm.nih.gov/18715936/

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,–

RPTEC/TERT1
Cat#: CHT-003-0002

Ordering now

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO

Related Products

ProxUp serum-free cell culture medium for growth of RPTEC/TERT1

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.

all about
our cells

read more

cisplatin-induced toxicity

RPTEC/TERT1 cells as relevant in vitro model to study kidney toxicity

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,–

RPTEC/TERT1
Cat#: CHT-003-0002

Ordering

ONLY FOR NON PROFIT

Ordering

FOR PROFIT INDUSTRY

Ordering

FOR PROFIT-CRO

Go to Top