CRL-2854 WPMY-1 人正常前列腺基质永生化细胞

ATCC® Number: CRL-2854™ Price:

Designations: WPMY-1

Depositors: MM Webber

Biosafety Level: 2 [Cells containing SV40 viral DNA sequence ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens （human）

Morphology: myofibroblast

CRL-2854 WPMY-1 人正常前列腺基质永生化细胞

Source: Organ: prostate

Tissue: stroma

Disease: normal

Cellular Products: fibronectin [89997]

smooth muscle alpha-actin

vimentin [89997]

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Isolation: Isolation date: 1992

Receptors: androgen receptor, expressed （ [89997] upregulated upon exposure to androgen）

Tumorigenic: No

Antigen Expression: kallikrein 3, KLK3 （prostate specific antigen, PSA）; Homo sapiens [89997]

Cytogenetic Analysis: At passage 66, a majority of the cells were in the 58-68 range; X, -Y. [89997]

Isoenzymes: AK-1, 1

ES-D, 2

G6PD, B

GLO-I, 1-2

Me-2, 0

PGM1, 2

PGM3, 1

Age: 54 years

Gender: male

Ethnicity: Caucasian, White

Comments: The myofibroblast stromal cell line, WPMY-1, was derived from stromal cells from the same peripheral zone of the histologically normal adult prostate, as that used for RWPE-1 cells （ATCC CRL-11609）. 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. Because of their derivation from the same peripheral zone of the prostate, the WPMY-1 cell line is especially useful for studies on paracrine and stromal : epithelial interactions.

The depositor reports that the RWPE-1 cell line （ATCC CRL-11609）, derived from the same prostate, was screened for Hepatitis B and C, and human immunodeficiency viruses, and was found to be negative.

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 5%.

Atmosphere: air, 95%; carbon dioxide （CO2）, 5%

Temperature: 37.0°C

Growth Conditions: Subculture cells before or upon reaching confluence. Do not allow cells to become super-confluent.

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 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 37C 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 approximay 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 4 X 10（3） to 6 X 10（3） viable cells/sq. cm is recommended.

7.Incubate cultures at 37C. We recommend that you maintain cultures at a cell concentraton between 6 X 10（3） and 5 X 10（4） cells/sq. cm.

Subc*tion Ratio: A subc*tion ratio of 1:3 to 1:6 is recommended

Medium Renewal: Every 48 hours

Preservation: Freeze medium: Complete growth medium supplemented with an additional 15% fetal bovine serum and 10% （v/v） DMSO

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 38 hours

Related Products: Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2002

recommended serum:ATCC 30-2020

derived from same individual:ATCC CRL-11609

derived from same individual:ATCC CRL-11610

derived from same individual:ATCC CRL-2849

derived from same individual:ATCC CRL-2850

derived from same individual:ATCC CRL-2852

derived from same individual:ATCC CRL-2851

References: 46793: Bello D, et al. Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215-1223, 1997. PubMed: 9214605

46795: 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

46799: 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

46950: 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

53180: 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

53181: 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

53182: 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

53183: 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

53192: 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

53193: 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

53194: Achanzar WE, et al. Cadmium-induced malignant transformation of human prostate epithelial cells. Cancer Res. 61: 455-458, 2001. PubMed: 11212230

53196: 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

53197: 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

89996: 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

89997: 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

89998: 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

90372: 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

90375: 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

90376: 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

CRL-2854 WPMY-1