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CP-D (CP-18821)


Product category
Human cells
Product type
hTERT-immortalized cell
Homo sapiens, human
Esophagus; Epithelium
Barretts Esophagus
3D cell culture
Drug development
High-throughput screening
Product format
Storage conditions
Vapor phase of liquid nitrogen
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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.

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.

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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


Specific applications
This well-characterized pre-malignant culture represents a unique tool for studying esophageal cancer progression.


Growth properties
The Barrett's esophagus cell line, CP-D (also identified as CP-18821) was derived from an endoscopic biopsy specimen obtained from a region of high-grade dysplasia. The cells were immortalized by transduction with a retroviral expression vector, pLXSN-hTERT, to create an immortalized cell line.
Immortalization method
hTERT expression
This is a hypotetraploid human cell line with the following derivative chromosomes consistently present at several different passages: add(2)(q13), der(3)t(3;8)(p10;q10), ider(7)(q10)dup(q31), der(12)t(12;13)(p10;q10), der(14)t(14;15)(q10;q10), der(15)t(15;22)(q10;q10), add(22)(q13)x2. In addition, there were consistent losses of one copy of chromosomes X, 10, 13, 14, 15, 19 and 20. Other less consistent structural aberrations were observed in some of the examined cells.
Antigen expression
Positive for epithelial marker pan-cytokeratin (immunocytochemistry)(verified at ATCC)
negative for gastric mucin (CLH2) (immunocytochemistry)(verified at ATCC)
Genes expressed
positive for epithelial marker pan-cytokeratin (immunocytochemistry), (verified at ATCC); negative for gastric mucin (CLH2) (immunocytochemistry), (verified at ATCC)

Terminal restriction fragment lengths (TRF) analyses show the cells have increased telomerase activity and extended telomeres of about 12 kb. Morphologically, the cell line is similar to early passage cultures exhibiting smaller cells with large nucleus to cytoplasm ratio. Genetic instability studies using flow cytometry and FISH reveal the retention of elevated tetraploidy (G2/tetraploidy) in the hTERT-immortalized cells, similar to the non-transduced parental cells.

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.

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
The base medium for this cell line is MCDB-153. To make the complete growth medium, add the following components to the base medium:
  • 0.4 µg/ml hydrocortisone
  • 20 ng/ml recombinant human EGF (Epidermal Growth Factor)
  • 8.4 µg/L cholera toxin
  • 20 mg/L adenine
  • 140 µg/ml BPE (Bovine Pituitary Extract)
  • 1x ITS Supplement (Sigma; I1884) [5 µg/ml Insulin; 5 µg /ml Transferrin; 5 ng/ml Sodium Selenite; final concs.]
  • 4 mM glutamine
  • fetal bovine serum to a final concentration of 5%
  • Note: To prepare Cholera toxin (Stock 100 µg/mL) : 0.5 mg Cholera toxin (Sigma C8052) + 5 mL dH20.  Aliquot into microcentrifuge tubes.   Add 84 µL of this 100 µg/mL stock solution to 1L of MCDB-153 base medium.

    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 phase 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. Add 2.0 to 3.0 mL of 0.25% (w/v) Trypsin - 0.53 mM 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.
    3. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
    4. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
    5. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 104 to 2 X 104 viable cells/cm2 is recommended.
    6. Incubate cultures at 37.0°C. Subculture when cells reach a concentration between 7 X 104 and 1 X 105 cells/cm2.
    Subcultivation ratio: A subcultivation ratio of 1:2 to 1:5 is recommended.
    Medium renewal: every 3 to 4 days
    Reagents for cryopreservation
    RPMI-1640 Medium supplemented with 10% (v/v) FBS and 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 39 hrs
    STR profiling
    Amelogenin: X,Y
    CSF1PO: 12
    D13S317: 12
    D16S539: 10,13
    D5S818: 9,12
    D7S820: 10,11
    TH01: 9,9.3
    TPOX: 8
    vWA: 16
    D3S1358: 15,16
    D21S11: 29,32.2
    D18S51: 13,16
    Penta_E: 10
    Penta_D: 12,13
    D8S1179: 11,15
    FGA: 20,22
    D19S433: 15,15.2
    D2S1338: 23,25


    B. Reid
    Year of origin

    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.

    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.


    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

    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.



    Curated Citations

    Palanca-Wessels MC, et al. Genetic Analysis of Long-term Barrett's Esophagus Epithelial Cultures Exhibiting Cytogenetic and Ploidy Abnormalities. Gastroentrology 114:114-295, 1998. PubMed: 9453489

    Palanca-Wessels MC, et al. Extended lifespan of Barrett's esophagus epithelium transduced with the human telomerase catalytic subunit: a useful in vitro model. Carcinogenesis 24(7): 1183-1190, 2003. PubMed: 12807723

    Barrett MT, et al. Molecular Phenotype of Spontaneously Arising 4N (G2-Tetraploid) Intermediates of Neoplastic Progression in Barrett's Esophagus. Cancer Res. 63: 4211-4217, 2003. PubMed: 12874028

    Maley CC, et al. Genetic clonal diversity predicts progression to esophageal adenocarcinoma. Nat. Genet. 38(4): 468-473, 2006. PubMed: 16565718

    Frequently Asked Questions

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