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Stromal cells were immortalized with SV40-large-T antigen gene, using a pRSTV plasmid construct. 

WPMY-1 stromal cells belong to a family of cell lines derived from the same prostate as the epithelial RWPE-1 cells and all of its epithelial derivatives.

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 of 0.025% Trypsin - 0.26 mM EDTA solution (1:1 dilution of 0.05% Trypsin - 0.53 mM EDTA in D-PBS) to the 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 6.0 to 8.0 mL of 0.1% Soybean Trypsin Inhibitor (or 2% fetal bovine serum in D-PBS) and aspirate cells by gently pipetting.
5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum between 1 x 10⁴ to 2 x 10⁴ viable cells/cm² is recommended.
7. Incubate cultures at 37°C. We recommend that you maintain cultures at a cell concentraton between 4 x 10⁴ and 8 x 10⁴ cells/cm².

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

at Passage 22 cells did not form tumors when injected subcutaneously

colony forming efficiency (CFE) of 0.7%

Mukta M Webber, personal communication

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