To ATCC Valued Customers,

ATCC stands ready to support our customers’ needs during the coronavirus pandemic. If you experience any issues with your products or services, please contact ATCC Customer Service at sales@atcc.org. For Technical questions please contact tech@atcc.org. Thank you.
X

RPTEC/TERT1 (ATCC® CRL-4031)

Organism: Homo sapiens, human  /  Cell Type: Epithelial cells immortalized with pLXSN-hTERT retroviral transfection  /  Tissue: Renal cortex; proximal tubules, epithelium  / 

Permits and Restrictions

View Permits View Restrictions

Organism Homo sapiens, human
Tissue Renal cortex; proximal tubules, epithelium
Cell Type Epithelial cells immortalized with pLXSN-hTERT retroviral transfection
Product Format frozen 1.0 mL
Morphology Epithelial-like
Culture Properties Adherent
Biosafety Level 2  [Cells contain SV40 viral DNA sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Age Adult
Gender Male
Applications
These cells are proposed to be a valuable model system not only for cell biology, but also toxicology, drug screening, biogerontology and tissue engineering.
Storage Conditions Liquid nitrogen vapor phase
Images
Antigen Expression Antigen expression: This cell line is positive for epithelial marker pan-cytokeratin (immunocytochemistry)(verified at ATCC), positive for epithelial cell adhesion molecule E-cadherin (immunocytochemistry) (verified at ATCC), 

The RPTEC/TERT1 cells express both Aminopeptidase N (verified at ATCC) and γ-Glutamyl Transferase (GGT) that are located in the brush border of the renal proximal tubular epithelium.
Comments

The RPTEC/TERT1 cells specifically respond to parathyroid hormone (PTH) but not arginine vasopressin (AVP), and react with enhanced ammonia genesis on lowering of the environmental pH.

The RPTEC/TERT1 cells exhibit sodium-dependent uptake of phosphate as well as intact functionality of the megalin/cubilin transport system.

RPTEC/TERT1 cells show the characteristic morphology and functional properties of normal proximal tubular epithelial cells.

When cultured on the Corning™ Transwell™ Permeable membrane cell culture insert, the RPTEC/TERT1 cells at confluence form intact functional barrier as indicated by stabilized Trans-Epithelial Electrical Resistance (TEER) across the membrane.

Complete Growth Medium

The base medium for this cell line is ATCC-formulated DMEM:F12 Medium (ATCC® 30-2006™).

To make the complete growth medium, add hTERT RPTEC Growth Kit (ATCC® ACS-4007™) to the base medium. The final concentration for each growth kit component in the complete hTERT immortalized RPTEC growth medium is as follows:  

  • 5 pM triiodo-L-thyronine
  • 10 ng/mL recombinant human EGF
  • 3.5 µg/mL ascorbic acid
  • 5.0 µg/mL human transferrin
  • 5.0 µg/mL insulin
  • 25 ng/mL prostaglandin E1
  • 25 ng/mL hydrocortisone
  • 8.65 ng/mL sodium selenite
  • 1.2 mg/mL sodium bicarbonate

Required but not supplied: G418 solution MUST be added to the above medium to a final concentration of 0.1 mg/mL G418 to maintain the selective pressure for immortalization

Note: Do not filter complete medium.

This medium is formulated for use with a 5% CO2 in air atmosphere.


Subculturing
Volumes are given for a 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Subculture when the culture is about 90% confluence. Expected cell yield is between 1.5 x 105 and 2 x 105 viable cells/cm2.
  2. Remove and discard culture medium.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA for Primary Cells (ATCC PCS-999-003) to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 3 to 8 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Place at 37°C to facilitate dispersal.
  4. To stop trypsinization, add 2.0 to 3.0 mL of 0.1% Soybean Trypsin Inhibitor and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a 15-mL centrifuge tube and spin at approximately 250 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 to 6 x 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C.
Subcultivation ratio: A subcultivation ratio of 1:3 to 1:4 is recommended.
Medium renewal: 2 to 3 times weekly
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Aminal Cells: A Manual of Basic Techniques by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
Cryopreservation
Freeze medium: Complete culture medium + 10% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Volume 1.0 mL
STR Profile
CSF1PO: 11
D13S317: 11, 13
D16S539: 11, 12
D5S818: 9, 11
D7S820: 10
TH01: 9, 9.3
TPOX: 8, 11
vWA: 16, 18
Amelogenin: XY
Population Doubling Level (PDL) As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation. In addition, it has been verified that no gross changes are observed in karyotype and morphology during the first 10 population doublings.
Name of Depositor R Grillari-Voglauer
Year of Origin July 2004
References

Wieser M, et al. hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics. Am. J. Physiol. Renal Physiol. 295: 1365-1375, 2008. PubMed: 18715936

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13.

Limonciel A, et al. Comparison of base-line and chemical-induced transcriptomic responses in HepaRG and RPTEC/TERT1 cells using TempO-Seq. Arch Toxicol 92(8):2517-2531, 2018. PubMed: 30008028

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

Aschauer L, et al. Expression of xenobiotic transporters in the human renal proximal tubule cell line RPTEC/TERT1. Toxicol In Vitro 30(1 Pt A):95-105, 2015. PubMed: 25500123

Simon BR, et al. The RPTEC/TERT1 cell line models key renal cell responses to the environmental toxicants, benzo[a]pyrene and cadmium. Toxicol Rep 1:231-242, 2014. PubMed: 25126521

Secker PF, et al. RPTEC/TERT1 cells form highly differentiated tubules when cultured in a 3D matrix. ALTEX 35(2):223-234, 2017. PubMed: 29197217

Shrestha S, et al. Human renal tubular cells contain CD24/CD133 progenitor cell populations: Implications for tubular regeneration after toxicant induced damage using cadmium as a model. Toxicol Appl Pharmacol 331:116-129, 2017. PubMed: 28587817

Simon BR, et al. Cadmium alters the formation of benzo[a]pyrene DNA adducts in the RPTEC/TERT1 human renal proximal tubule epithelial cell line. Toxicol Rep 1:391-400, 2014. PubMed: 25170436

Soodvilai S, et al. Interaction of pharmaceutical excipients with organic cation transporters. Int J Pharm 520(1-2):14-20, 2017. PubMed: 28131852

Shah H, et al. Gene expression study of phase I and II metabolizing enzymes in RPTEC/TERT1 cell line: application in in vitro nephrotoxicity prediction. Xenobiotica 47(10):837-843, 2017. PubMed: 27616666

Kramer NI, et al. Biokinetics in repeated-dosing in vitro drug toxicity studies. Toxicol In Vitro 30(1 Pt A):217-24, 2015. PubMed: 26362508

Aschauer L, et al. Application of RPTEC/TERT1 cells for investigation of repeat dose nephrotoxicity: A transcriptomic study. Toxicol In Vitro 30(1 Pt A):106-16, 2015. PubMed: 25450743

Wilmes A, et al. Mechanism of cisplatin proximal tubule toxicity revealed by integrating transcriptomics, proteomics, metabolomics and biokinetics. Toxicol in Vitro 30(1 Pt A):117-27, 2015. PubMed: 25450742

Wilmes A, et al. Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress. J Proteomics V79:180-194, 2013. PubMed: 23238060

Wilmes A, et al. Evidence for a role of claudin 2 as a proximal tubular stress responsive paracellular water channel. Toxicol Appl Pharmacol 279(2):163-72, 2014. PubMed: 24907557

Jennings P, et al. Interleukin-19 as a translational indicator of renal injury. Arch Toxicol 89(1):101-6, 2015. PubMed: 24714768

Ranninger C, et al. Nephron Toxicity Profiling via Untargeted Metabolome Analysis Employing a High Performance Liquid Chromatography-Mass Spectrometry-based Experimental and Computational Pipeline. J Biol Chem 290(31): 19121-32, 2015 PubMed: 26055719

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

For commercial accounts, this cell line is only distributed under the terms of a fully signed and executed ATCC® Material Transfer Agreement and Addendum. If the commercial account is screening per completed Addendum, the recipient will be required to pay a Screening Fee (ATCC® ACS-2103F™).

Screening Use is defined as use of Biological Material in small molecule and biologic drug discovery, including initial target identification and validation, assay development, high throughput screening, hit identification, lead optimization, and selection of candidates for clinical development.

If the commercial account is not screening per the completed Addendum, the recipient will not be required to pay a Screening Fee.

References

Wieser M, et al. hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics. Am. J. Physiol. Renal Physiol. 295: 1365-1375, 2008. PubMed: 18715936

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13.

Limonciel A, et al. Comparison of base-line and chemical-induced transcriptomic responses in HepaRG and RPTEC/TERT1 cells using TempO-Seq. Arch Toxicol 92(8):2517-2531, 2018. PubMed: 30008028

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

Aschauer L, et al. Expression of xenobiotic transporters in the human renal proximal tubule cell line RPTEC/TERT1. Toxicol In Vitro 30(1 Pt A):95-105, 2015. PubMed: 25500123

Simon BR, et al. The RPTEC/TERT1 cell line models key renal cell responses to the environmental toxicants, benzo[a]pyrene and cadmium. Toxicol Rep 1:231-242, 2014. PubMed: 25126521

Secker PF, et al. RPTEC/TERT1 cells form highly differentiated tubules when cultured in a 3D matrix. ALTEX 35(2):223-234, 2017. PubMed: 29197217

Shrestha S, et al. Human renal tubular cells contain CD24/CD133 progenitor cell populations: Implications for tubular regeneration after toxicant induced damage using cadmium as a model. Toxicol Appl Pharmacol 331:116-129, 2017. PubMed: 28587817

Simon BR, et al. Cadmium alters the formation of benzo[a]pyrene DNA adducts in the RPTEC/TERT1 human renal proximal tubule epithelial cell line. Toxicol Rep 1:391-400, 2014. PubMed: 25170436

Soodvilai S, et al. Interaction of pharmaceutical excipients with organic cation transporters. Int J Pharm 520(1-2):14-20, 2017. PubMed: 28131852

Shah H, et al. Gene expression study of phase I and II metabolizing enzymes in RPTEC/TERT1 cell line: application in in vitro nephrotoxicity prediction. Xenobiotica 47(10):837-843, 2017. PubMed: 27616666

Kramer NI, et al. Biokinetics in repeated-dosing in vitro drug toxicity studies. Toxicol In Vitro 30(1 Pt A):217-24, 2015. PubMed: 26362508

Aschauer L, et al. Application of RPTEC/TERT1 cells for investigation of repeat dose nephrotoxicity: A transcriptomic study. Toxicol In Vitro 30(1 Pt A):106-16, 2015. PubMed: 25450743

Wilmes A, et al. Mechanism of cisplatin proximal tubule toxicity revealed by integrating transcriptomics, proteomics, metabolomics and biokinetics. Toxicol in Vitro 30(1 Pt A):117-27, 2015. PubMed: 25450742

Wilmes A, et al. Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress. J Proteomics V79:180-194, 2013. PubMed: 23238060

Wilmes A, et al. Evidence for a role of claudin 2 as a proximal tubular stress responsive paracellular water channel. Toxicol Appl Pharmacol 279(2):163-72, 2014. PubMed: 24907557

Jennings P, et al. Interleukin-19 as a translational indicator of renal injury. Arch Toxicol 89(1):101-6, 2015. PubMed: 24714768

Ranninger C, et al. Nephron Toxicity Profiling via Untargeted Metabolome Analysis Employing a High Performance Liquid Chromatography-Mass Spectrometry-based Experimental and Computational Pipeline. J Biol Chem 290(31): 19121-32, 2015 PubMed: 26055719