WPE-stem (ATCC® CRL-2887)

Organism: Homo sapiens, human  /  Cell Type: epithelial  /  Tissue: prostate, normal, peripheral zone  /  Disease: Papilloma

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Organism Homo sapiens, human
Tissue prostate, normal, peripheral zone
Cell Type epithelial
Product Format frozen
Morphology epithelial
Culture Properties loosely adherent
Biosafety Level 2 [cells contain human HPV-18 viral DNA sequences]
Disease Papilloma
Age 54
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Images
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]
Clinical Data
54
Caucasian
male
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
Receptor Expression
androgen receptor expressed, slightly upregulated upon exposure to androgen 
Tumorigenic No
Effects
No, the cells are not tumorigenic in nude mice
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).

Complete Growth 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.
  • Subculturing

    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
    Cryopreservation
    Freeze medium: Complete growth medium described above supplemented with 15% fetal bovine serum and 10% (v/v) DMSO. Cell culture tested DMSO is available as ATCC® Catalog No. 4-X.
    Storage temperature: liquid nitrogen vapor phase
    Culture Conditions
    Atmosphere: air, 95%; carbon dioxide (CO2), 5%
    Temperature: 37°C
    Growth Conditions: 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 microgram each/cm2.
    STR Profile
    Amelogenin: X,Y
    CSF1PO: 13
    D13S317: 8,14
    D16S539: 9,11
    D5S818: 12,15
    D7S820: 10,11
    THO1: 8
    TPOX: 8,11
    vWA: 14,18
    Population Doubling Time about 24 hours
    Name of Depositor MM Webber, EJ Tokar
    Year of Origin 2004 [PubMed: 16351690]
    References

    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

    Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications Part 2. Tumorigenic cell lines. Prostate 30: 58-64, 1997. PubMed: 9018337

    Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications. Part 3. Oncogenes, suppressor genes, and applications. Prostate 30: 136-142, 1997. PubMed: 9051152

    Kremer R, et al. ras Activation of human prostate epithelial cells induces overexpression of parathyroid hormone-related peptide. Clin. Cancer Res. 3: 855-859, 1997. PubMed: 9815759

    Jacob K, et al. Osteonectin promotes prostate cancer cell migration and invasion: a possible mechanism for metastasis to bone. Cancer Res. 59: 4453-4457, 1999. PubMed: 10485497

    Achanzar WE, et al. Cadmium induces c-myc, p53, and c-jun expression in normal human prostate epithelial cells as a prelude to apoptosis. Toxicol. Appl. Pharmacol. 164: 291-300, 2000. PubMed: 10799339

    Achanzar WE, et al. Cadmium-induced malignant transformation of human prostate epithelial cells. Cancer Res. 61: 455-458, 2001. PubMed: 11212230

    Bello-DeOcampo D, et al. Laminin-1 and alpha6beta1 integrin regulate acinar morphogenesis of normal and malignant human prostate epithelial cells. Prostate 46: 142-153, 2001. PubMed: 11170142

    Webber MM, et al. Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 47: 1-13, 2001. PubMed: 11304724

    Quader ST, et al. Evaluation of the chemopreventive potential of retinoids using a novel in vitro human prostate carcinogenesis model. Mutat. Res. 496: 153-161, 2001. PubMed: 11551491

    Webber MM, et al. A human prostatic stromal myofibroblast cell line WPMY-1: a model for stromal-epithelial interactions in prostatic neoplasia. Carcinogenesis 20: 1185-1192, 1999. PubMed: 10383888

    Bello-DeOcampo D, et al. The role of alpha 6 beta 1 integrin and EGF in normal and malignant acinar morphogenesis of human prostatic epithelial cells. Mutat. Res. 480-481: 209-217, 2001. PubMed: 11506815

    Webber MM, et al. Modulation of the malignant phenotype of human prostate cancer cells by N-(4-hydroxyphenyl)retinamide (4-HPR). Clin. Exp. Metastasis 17: 255-263, 1999. PubMed: 10432011

    Sharp RM, et al. N-(4-hydroxyphenyl)retinamide (4-HPR) decreases neoplastic properties of human prostate cells: an agent for prevention. Mutat. Res. 496: 163-170, 2001. PubMed: 11551492

    Carruba G, et al. Regulation of cell-to-cell communication in non-tumorigenic and malignant human prostate epithelial cells. Prostate 50: 73-82, 2002. PubMed: 11816015

    Achanzar WE, et al. Altered apoptotic gene expression and acquired apoptotic resistance in cadmium-transformed human prostate epithelial cells. Prostate 52: 236-244, 2002. PubMed: 12111698

    Carruba G, et al. Intercellular communication and human prostate carcinogenesis. Ann. N.Y. Acad. Sci. 963: 156-168, 2002. PubMed: 12095941

    Saladino F, et al. Connexin expression in nonneoplastic human prostate epithelial cells. Ann. N.Y. Acad. Sci. 963: 213-217, 2002. PubMed: 12095946

    Hegarty PK, et al. Effects of cyclic stretch on prostatic cells in culture. J. Urol. 168: 2291-2295, 2002. PubMed: 12394777

    Lugassy C, et al. Human melanoma cell migration along capillary-like structures in vitro: a new dynamic model for studying extravascular migratory metastasis. J. Invest. Dermatol. 119: 703-704, 2002. PubMed: 12230517

    Achanzar WE, et al. Inorganic arsenite-induced malignant transformation of human prostate epithelial cells. J. Natl. Cancer Inst. 94: 1888-1891, 2002. PubMed: 12488483

    Tokar EJ, et al. Stem/progenitor and intermediate cell types and the origin of human prostate cancer. Differentiation 73: 463-473, 2005. PubMed: 16351690

    Achanzar WE, et al. Human prostate cell lines mimic heterogeneity of cadhedrin expression in human prostate cancer. Urol. Oncol. 4: 15-25, 2004.

    Bramblia EM, et al. Chronic arsenic-exposed human prostate epithelial cells exhibit stable arsenic tolerance: mechanistic implications of altered cellular glutathione and glutathione S-transferase. Toxicol. Appl. Pharmacol. 183: 99-107, 2002. PubMed:12387749.

    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
    References

    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

    Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications Part 2. Tumorigenic cell lines. Prostate 30: 58-64, 1997. PubMed: 9018337

    Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications. Part 3. Oncogenes, suppressor genes, and applications. Prostate 30: 136-142, 1997. PubMed: 9051152

    Kremer R, et al. ras Activation of human prostate epithelial cells induces overexpression of parathyroid hormone-related peptide. Clin. Cancer Res. 3: 855-859, 1997. PubMed: 9815759

    Jacob K, et al. Osteonectin promotes prostate cancer cell migration and invasion: a possible mechanism for metastasis to bone. Cancer Res. 59: 4453-4457, 1999. PubMed: 10485497

    Achanzar WE, et al. Cadmium induces c-myc, p53, and c-jun expression in normal human prostate epithelial cells as a prelude to apoptosis. Toxicol. Appl. Pharmacol. 164: 291-300, 2000. PubMed: 10799339

    Achanzar WE, et al. Cadmium-induced malignant transformation of human prostate epithelial cells. Cancer Res. 61: 455-458, 2001. PubMed: 11212230

    Bello-DeOcampo D, et al. Laminin-1 and alpha6beta1 integrin regulate acinar morphogenesis of normal and malignant human prostate epithelial cells. Prostate 46: 142-153, 2001. PubMed: 11170142

    Webber MM, et al. Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 47: 1-13, 2001. PubMed: 11304724

    Quader ST, et al. Evaluation of the chemopreventive potential of retinoids using a novel in vitro human prostate carcinogenesis model. Mutat. Res. 496: 153-161, 2001. PubMed: 11551491

    Webber MM, et al. A human prostatic stromal myofibroblast cell line WPMY-1: a model for stromal-epithelial interactions in prostatic neoplasia. Carcinogenesis 20: 1185-1192, 1999. PubMed: 10383888

    Bello-DeOcampo D, et al. The role of alpha 6 beta 1 integrin and EGF in normal and malignant acinar morphogenesis of human prostatic epithelial cells. Mutat. Res. 480-481: 209-217, 2001. PubMed: 11506815

    Webber MM, et al. Modulation of the malignant phenotype of human prostate cancer cells by N-(4-hydroxyphenyl)retinamide (4-HPR). Clin. Exp. Metastasis 17: 255-263, 1999. PubMed: 10432011

    Sharp RM, et al. N-(4-hydroxyphenyl)retinamide (4-HPR) decreases neoplastic properties of human prostate cells: an agent for prevention. Mutat. Res. 496: 163-170, 2001. PubMed: 11551492

    Carruba G, et al. Regulation of cell-to-cell communication in non-tumorigenic and malignant human prostate epithelial cells. Prostate 50: 73-82, 2002. PubMed: 11816015

    Achanzar WE, et al. Altered apoptotic gene expression and acquired apoptotic resistance in cadmium-transformed human prostate epithelial cells. Prostate 52: 236-244, 2002. PubMed: 12111698

    Carruba G, et al. Intercellular communication and human prostate carcinogenesis. Ann. N.Y. Acad. Sci. 963: 156-168, 2002. PubMed: 12095941

    Saladino F, et al. Connexin expression in nonneoplastic human prostate epithelial cells. Ann. N.Y. Acad. Sci. 963: 213-217, 2002. PubMed: 12095946

    Hegarty PK, et al. Effects of cyclic stretch on prostatic cells in culture. J. Urol. 168: 2291-2295, 2002. PubMed: 12394777

    Lugassy C, et al. Human melanoma cell migration along capillary-like structures in vitro: a new dynamic model for studying extravascular migratory metastasis. J. Invest. Dermatol. 119: 703-704, 2002. PubMed: 12230517

    Achanzar WE, et al. Inorganic arsenite-induced malignant transformation of human prostate epithelial cells. J. Natl. Cancer Inst. 94: 1888-1891, 2002. PubMed: 12488483

    Tokar EJ, et al. Stem/progenitor and intermediate cell types and the origin of human prostate cancer. Differentiation 73: 463-473, 2005. PubMed: 16351690

    Achanzar WE, et al. Human prostate cell lines mimic heterogeneity of cadhedrin expression in human prostate cancer. Urol. Oncol. 4: 15-25, 2004.

    Bramblia EM, et al. Chronic arsenic-exposed human prostate epithelial cells exhibit stable arsenic tolerance: mechanistic implications of altered cellular glutathione and glutathione S-transferase. Toxicol. Appl. Pharmacol. 183: 99-107, 2002. PubMed:12387749.