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WPE-stem

CRL-2887

Product category
Human cells
Organism
Homo sapiens, human
Cell type
epithelial cell
Morphology
epithelial
Tissue
Prostate; Peripheral zone
Disease
Papilloma
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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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 18 (HPV-18) 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
Loosely adherent
Derivation
WPE-stem cells were derived from the RWPE-1 cell line (ATCC CRL-11609) after two consecutive cycles of single cell cloning. To establish the RWPE-1 cell line, epithelial cells from the peripheral zone of a histologically normal adult human prostate were transfected with a plasmid carrying one copy of the human papilloma virus 18 (HPV-18) genome. The WPE-int cell line (ATCC CRL-2888), which has an intermediate phenotype on the path to luminal cell differentiation, was also derived from RWPE-1 cells after single cell cloning. [PubMed: 16351690]
Age
54 years
Ethnicity
White
Gender
Male
Tumorigenic
No;
No, the cells are not tumorigenic in nude mice
Antigen expression
Weak expression of prostate-specific antigen (PSA) and androgen receptor (AR) upon exposure to androgen. RefTokar EJ, et al. Stem/progenitor and intermediate cell types and the origin of human prostate cancer. Differentiation 73: 463-473, 2005. PubMed: 16351690
Expression markers
Androgen receptor, expressed, slightly upregulated upon exposure to androgen
Comments

WPE-stem cells are loosely adherent and exhibit features characteristic of stem/progenitor cells present in the embryonic urogenital sinus and in adult prostatic epithelium, including p63 and ABCG2.

The cells show high expression of cytokeratin 5 and 14 and MMP-2 but low expression of cytokeratin 18.

They are androgen-independent for growth and survival.

Cells grow in soft agar with a cloning efficiency of 0.9%.

Cells are not tumorigenic in nude mice even at 6 months after injection.

WPE-stem cells were screened for CMV, HBV, HCV, HTLV 1, HTLV 2, HIV 1, HIV 2, JCV, and MoMuLV DNA sequences. Cells were also tested for 25 species of mycoplasma and Acholeplasma laidlawii . Cells tested negative for all of the above. (personal communication from depositor).

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 provided by Invitrogen (GIBCO) as part of a kit: Keratinocyte Serum Free Medium (K-SFM), Kit Catalog Number 17005-042. This kit is supplied with each of the two additives required to grow this cell line (bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF). To make the complete growth medium, you will need to add the following components to the base medium:
  • 0.05 mg/ml BPE - provided with the K-SFM kit
  • 5 ng/ml EGF - provided with the K-SFM kit. NOTE: Do not filter complete medium.
  • Temperature
    37°C
    Atmosphere
    95% Air, 5% CO2
    Handling procedure

    To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

    Note: Culture flasks should be pre-coated with a mixture of Mouse Collagen Type IV and Human Fibronectin (Sigma Cat. No. F-0895) 2.5µg each/ cm2.
    1. 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).
    2. 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 of the operations from this point on should be carried out under strict aseptic conditions.
    3. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
    4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a pre-coated 25 cm2 or a 75 cm2 culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.4).
    5. Incubate the culture at 37°C in a suitable, humidified incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product.

     

    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. Subculture cells before they reach confluence. Do not allow cells to become confluent.


    Note: Culture flasks should be pre-coated with a mixture of Mouse Collagen Type IV and Human Fibronectin (Sigma Cat. No. F-0895) 2.5 µg each/cm2.
    1. Collect spent medium containing viable, floating, single cells, and prostaspheres.
    2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).
    3. 3.0 to 4.0 mL of 0.05% Trypsin - 0.53 mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37°C incubator for 3 to 5 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 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.
    4. Add 6.0 to 8.0 mL of 0.1% Soybean Trypsin Inhibitor, or 2% fetal bovine serum in D-PBS, as appropriate, add the spent medium containing floating cells, and aspirate cells by gently pipetting.
    5. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 7 minutes.
    6. Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new pre-coated culture flasks. An inoculum of 2 X 103 to 4 X 103 viable cells/cm2 is recommended.
    7. Place culture vessels in incubators at 37°C. Cells grown under serum-free or reduced serum conditions may not attach strongly during the 24 hours after subculture and should be disturbed as little as possible during that period.

    Interval: Subculture cells before they reach confluence. Do not allow cells to become confluent. Subculture when cell concentration is between 1 X 105 and 1.5 X 105 cells/cm2.
    Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
    Medium Renewal: Every 48 hours
    Reagents for cryopreservation
    Complete growth medium supplemented with 15% (v/v) fetal bovine serum and 10% DMSO (ATCC 4-X)

    Quality control specifications

    Mycoplasma contamination
    Not detected
    Population doubling time
    Approximately 24 hrs
    STR profiling
    Amelogenin: X,Y
    CSF1PO: 13
    D13S317: 8,14
    D16S539: 9,11
    D5S818: 12,15
    D7S820: 10,11
    TH01: 8
    TPOX: 8,11
    vWA: 14,18
    D3S1358: 15,16
    D21S11: 29,31
    D18S51: 14,16
    Penta_E: 5,12
    Penta_D: 10,13
    D8S1179: 10,14
    FGA: 25
    D19S433: 13
    D2S1338: 17,20

    History

    Depositors
    MM Webber, EJ Tokar
    Year of origin
    2004

    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

    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

    Bello D, et al. Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215-1223, 1997. PubMed: 9214605

    Webber MM, et al. Acinar differentiation by non-malignant immortalized human prostatic epithelial cells and its loss by malignant cells. Carcinogenesis 18: 1225-1231, 1997. PubMed: 9214606

    Webber MM, et al. Prostate specific antigen and androgen receptor induction and characterization of an immortalized adult human prostatic epithelial cell line. Carcinogenesis 17: 1641-1646, 1996. PubMed: 8761420

    Okamoto M, et al. Interleukin-6 and epidermal growth factor promote anchorage-independent growth of immortalized human prostatic epithelial cells treated with N-methyl-N-nitrosourea. Prostate 35: 255-262, 1998. PubMed: 9609548

    Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications. Part I. Cell markers and immortalized nontumorigenic cell lines. Prostate 29: 386-394, 1996. PubMed: 8977636

    View All Curated Citations for this Product

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