American Type Culture Collection (ATCC) Logo American Type Culture Collection (ATCC) Logo 0
  • Quick Order
  • Careers
  • Support

hTERT-HPNE E6/E7/st

CRL-4037


Use promo code ATCC-000032 at checkout to save 10% on this product.*

*This offer is effective until December 31, 2024. The 10% discount applies only to the following products: CRL-3290™, CRL-3397™, CRL-4000™, CRL-4001™, CRL-4005™, CRL-4031™, CRL-4037™, CRL-4051™, SCRC-4000™, SCRC-4001™; all other items are excluded from this promotion. For non-profit customers, the promotional discount is applied to the customer-specific pricing. The promo code must be applied at the time of checkout. The promo code must be applied in the “Promotional code” field in the shopping cart. Void where prohibited. Restrictions may apply.

Product category
Human cells
Product type
hTERT-immortalized cell
Organism
Homo sapiens, human
Morphology
epithelial-like
Tissue
Pancreas; Duct
Applications
3D cell culture
Drug development
High-throughput screening
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
Buy Now
Price: $6,256.00 EA
Discounts may be available for our fellow nonprofit organizations. Login to see your price.

Generally ships within 1-3 business days

Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Cells contain Human papillomavirus type 16 (HPV-16) sequences

Cells contain SV40 sequences

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

Characteristics

Growth properties
Adherent
Derivation
Cells were derived by infection of hTERT-HPNE E6/E7 cells (ATCC CRL-4036) with retroviral vector (pBabeZeo) carrying the SV40 small t antigen
Age
52 years
Gender
Male
Immortalization method
hTERT expression
Karyotype
This is a pseudodiploid human cell line of male origin with a der(21)t(17;21)(q21.3;p13). Another subclone, present at earlier passages, may contain the additional derivative chromosome: der(3)t(3;18)(p21.1;q11.2). Overall, the cell line has a relatively stable karyotype.
Tumorigenic
No
Antigen expression
Positive for nestin (flow cytometry) (verified at ATCC)
Genes expressed
positive for nestin (flow cytometry), (verified at ATCC)

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium

Low Glucose DMEM + 25% INCELL M3:Base F + 5% FBS + 10 ng/mL human rEGF + 5.5 mM D-glucose + 2mM L-glutamine + 750 ng/mL Puromycin

  • 375 mL Low Glucose DMEM (Sigma catalog # D5921)
  • 125 mL INCELL M3: Base F (Fisher Scientific catalog # M300F-500)
  • 27 mL FBS (ATCC catalog # 30-2020)
  • 5.4 mL hr EGF Stock (1 µg/mL, Gibco catalog # PHG0314)
  • 5.4 mL L-glutamine (ATCC catalog # 30-2214)
  • 1.4 mL D-glucose (Sigma catalog # G8644)
  • 1 mL Puromycin Stock (405 µg/mL, Gibco catalog # A11138-03)
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If storage of the frozen culture is necessary upon arrival, store the vial in liquid nitrogen vapor and NOT at -70°C.  Storage at -70°C will result in loss of viability.

  1. Prepare a 25-cm2 or a 75-cm2 culture flask containing the recommended complete culture medium. Prior to the addition of the vial contents, the vessel containing the growth medium should be placed in the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6) and to avoid excessive alkalinity of the medium during recovery of the cells.
  2. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  3. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All operations from this point on should be carried out under strict aseptic conditions.
  4. Transfer the vial contents to a centrifuge tube containing 9.0 ml of complete culture medium and centrifuge the cell suspension at approximately 125 x g for 5 to 7 minutes. 
  5. Discard the supernatant and resuspend the cells in fresh growth medium (see the batch-specific information for the recommended dilution ratio). Add this suspension to the prepared culture vessel.
  6. Incubate the culture at 37°C in a suitable incubator. 
  7. A 5% CO2/95% air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 2.0 to 3.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 4 X 103 to 6 X 103 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C. Subculture when cell density reaches between 5 X 104 and 6 X 104 cells/cm2.
Subcultivation ratio: 1:8 to 1:12 twice weekly
Medium renewal: every 2 to 3 days
Reagents for cryopreservation
Fetal bovine serum supplemented with 10% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected
Population doubling capacity
≥ 15 in complete growth medium
Population doubling time
Approximately 29 hrs
STR profiling
Amelogenin: X,Y
CSF1PO: 12
D13S317: 12,13
D16S539: 12,13
D5S818: 11
D7S820: 9,10
TH01: 8,9
TPOX: 8,11
vWA: 17
D3S1358: 15
D21S11: 30
D18S51: 13,16
Penta_E: 7,10
Penta_D: 11
D8S1179: 10,13
FGA: 23,24
D19S433: 12,16.2
D2S1338: 17,22

History

Depositors
M Ouellette
Year of origin
2002

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Material Transfer Agreement Addendum for Screening Applications

For-profit organizations
For every order of this item, you must provide a signed Material Transfer Agreement Addendum for Screening Applications. We cannot ship this item until we receive this addendum. The person signing the addendum as the principal investigator must match the end user as listed on the applicable sales order for the item.

Email the signed addendum to [email protected] with a reference to both your account and sales order numbers. Once received, your addendum will be reviewed, and this item will be released for shipment if all requirements are met. Additional fees may apply if this product is being used for a screening use (ATCC ACS-2103F), and these fees will be applied after your order is confirmed. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Lee KM. et al. Notch2-positive progenitors with the intrinsic ability to give rise to pancreatic ductal cells. Lab. Invest. 85 (8): 1003-1012, 2005. Pubmed: 15924149

Lee KM. et al. Immortalization with telomerase of the Nestin-positive cells of the human pancreas. Biochem. Biophys. Res. Commun. 301(4):1038-1044, 2003. Pubmed: 12589817

Campbell PM, et al. K-Ras promotes growth transformation and invasion of immortalized human pancreatic cells by Raf and phosphatidylinositol 3-kinase signaling. Cancer Res. 67(5): 2098-2106, 2007. PubMed: 17332339

Campbell PM, et al. Ras-driven transformation of human nestin-positive pancreatic epithelial cells. Methods Enzymol. 439: 451-465, 2008. PubMed: 18374182

For product-related inquiries and issues, contact Technical Service:

Message Us

Hours of Operation

Monday - Friday
9:00am - 5:00pm
US Eastern Time