RWPE2-W99 (ATCC® CRL-2853)

Organism: Homo sapiens, human  /  Cell Type: epithelial  / 

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Organism Homo sapiens, human
Cell Type epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 cells containing human HPV-18 viral DNA sequences
Age 54 years
Gender male
Ethnicity Caucasian, White
Applications
The RWPE2-W99 cell line was derived, in 1999, from the RWPE-2 (ATCC CRL-11610) cell line by cloning in soft agar to select cells that show high expression of Ki-ras.
Storage Conditions liquid nitrogen vapor phase
Karyotype The depositor reports that at passage 44, a majority of the cells had a near diploid chromosome number of 48; X, -Y.
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Derivation
The RWPE2-W99 cell line was derived, in 1999, from the RWPE-2 (ATCC CRL-11610) cell line by cloning in soft agar to select cells that show high expression of Ki-ras. RWPE2-W99 cells show strong expression of Ki-ras and have characteristics similar to those of RWPE-2 cells. RWPE-2 cells were derived from RWPE-1 cells (ATCC CRL-11609) by transformation with Ki-ras using the Kirsten murine sarcoma virus (Ki-MuSV) to establish the RWPE-2 cells (PubMed: 9214605). 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 to establish the RWPE-1 cell line (ATCC CRL-11609). A family of tumorigenic cell lines that mimics multiple steps in prostate cancer progression was also derived from RWPE-1 cells by exposure to N-methyl-N-nitrosourea (MNU): WPE1-NA22 (ATCC CRL-2849), WPE1-NB14 (ATCC CRL-2850), WPE1-NB11 (ATCC CRL-2851) and WPE1-NB26 (ATCC CRL-2852).The depositor reports that this cell line was screened for Hepatitis B and C, and human immunodeficiency viruses, and was found to be negative.
Clinical Data
male
Caucasian, White
54 years
Antigen Expression
kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens, expressed (upon exposure to androgen)
Receptor Expression
androgen receptor, expressed
Genes Expressed
WPE1-NB26 cells were derived from RWPE-1 cells (ATCC CRL-11609) after exposure to N-methyl-N-nitrosourea (MNU) [PubMed: 11304724].
cytokeratin 18,cytokeratin 8,kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens, expressed (upon exposure to androgen),The RWPE2-W99 cell line was derived, in 1999, from the RWPE-2 (ATCC CRL-11610) cell line by cloning in soft agar to select cells that show high expression of Ki-ras.
Cellular Products
cytokeratin 18
cytokeratin 8
Tumorigenic Yes
Effects
Yes, in nude mice
Yes, the cells form colonies in soft agar
Comments
The RWPE2-W99 cell line was derived, in 1999, from the RWPE-2 (ATCC CRL-11610) cell line by cloning in soft agar to select cells that show high expression of Ki-ras. RWPE2-W99 cells show strong expression of Ki-ras and have characteristics similar to those of RWPE-2 cells. RWPE-2 cells were derived from RWPE-1 cells (ATCC CRL-11609) by transformation with Ki-ras using the Kirsten murine sarcoma virus (Ki-MuSV) to establish the RWPE-2 cells (PubMed: 9214605). 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 to establish the RWPE-1 cell line (ATCC CRL-11609). A family of tumorigenic cell lines that mimics multiple steps in prostate cancer progression was also derived from RWPE-1 cells by exposure to N-methyl-N-nitrosourea (MNU): WPE1-NA22 (ATCC CRL-2849), WPE1-NB14 (ATCC CRL-2850), WPE1-NB11 (ATCC CRL-2851) and WPE1-NB26 (ATCC CRL-2852).
The depositor reports that this cell line was screened for Hepatitis B and C, and human immunodeficiency viruses, and was found to be negative.
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
    Protocol:
    1. Remove and discard culture medium.
    2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).
    3. Add 2.0 to 3.0 ml (to a T-25 flask) or 3.0 to 4.0 ml (to a T-75 flask) of 0.05% Trypsin - 0.53 mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37C incubator for 5 to 8 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 10 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, 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 culture vessels. An inoculum between 2 X 10(4) to 4 X 10(4) viable cells/sq. cm is recommended.
    7. Incubate cultures at 37C. We recommend that you maintain cultures at a cell concentration between 4 X 10(4) and 8 X 10(4) cells/sq. cm.
    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.
    Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
    Medium Renewal: Every 48 hours
    Cryopreservation
    Freeze medium: Complete growth medium supplemented with 15% fetal bovine serum and 10% (v/v) DMSO
    Storage temperature: liquid nitrogen vapor phase
    Culture Conditions
    Atmosphere: air, 95%; carbon dioxide (CO2), 5%
    Temperature: 37.0°C
    Growth Conditions: Subculture cells before they reach confluence. Do not allow cells to become confluent.
    STR Profile
    Amelogenin: X,Y
    CSF1PO: 13
    D13S317: 8,14
    D16S539: 9,11
    D5S818: 12,15
    D7S820: 10,11
    THO1: 8,9.3
    TPOX: 8,11
    vWA: 14,18
    Population Doubling Time 36 hours
    Name of Depositor MM Webber
    Year of Origin 1999
    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

    upon exposure to androgen

    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

    upregulated upon exposure to androgen

    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

    Brambila 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

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

    RWPE2-W99 cells form small tumors when injected subcutaneously, with Matrigel, in nude mice.

    small colonies are formed

    Mukta M Webber, personal communication

    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

    upon exposure to androgen

    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

    upregulated upon exposure to androgen

    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

    Brambila 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

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

    RWPE2-W99 cells form small tumors when injected subcutaneously, with Matrigel, in nude mice.

    small colonies are formed

    Mukta M Webber, personal communication