RWPE2-W99細(xì)胞

細(xì)胞形態(tài)： 上皮樣

器官來源： 前列腺

細(xì)胞類型： 上皮細(xì)胞

ATCC Number： CRL-2853?

相關(guān)**： 正常

是否是腫瘤細(xì)胞： 0

物種來源： 人

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

運(yùn)輸方式： 凍存運(yùn)輸

年限： 54 years

數(shù)量： 大量

Designations: RWPE2-W99

RWPE2-W99細(xì)胞Depositors: MM Webber

Biosafety Level: 2 [cells containing human HPV-18 viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial

Source: Organ: prostate

Disease: normal

Cell Type: epithelial

Cellular Products: cytokeratin 18

cytokeratin 8

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Isolation: RWPE2-W99細(xì)胞Isolation date: 1999

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.

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

Tumorigenic: Yes

Antigen Expression: kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens, expressed (upon exposure to androgen) (upon exposure to androgen)

DNA Profile (STR): 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

Cytogenetic Analysis: The depositor reports that at passage 44, a majority of the cells had a near diploid chromosome number of 48; X, -Y.

Age: 54 years

Gender: male

Ethnicity: RWPE2-W99細(xì)胞Caucasian, White

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 ***** 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.

Propagation: ATCC 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.

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.

Subculturing: Protocol:

Remove and discard culture medium.

Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).

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: RWPE2-W99細(xì)胞To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.

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.

Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 7 minutes.

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.

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

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

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 36 hours

Related Products: derived from same individual:ATCC CRL-11609

parental cell line: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 ***** 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 ***** 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 ***** 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 ***** 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 ***** 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

90377: 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

90378: Carruba G, et al. Intercellular communication and human prostate carcinogenesis. Ann. N.Y. Acad. Sci. 963: 156-168, 2002. PubMed: 12095941

90379: Saladino F, et al. Connexin expression in nonneoplastic human prostate epithelial cells. Ann. N.Y. Acad. Sci. 963: 213-217, 2002. PubMed: 12095946

90380: Hegarty PK, et al. Effects of cyclic stretch on prostatic cells in culture. J. Urol. 168: 2291-2295, 2002. PubMed: 12394777

90383: 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

90384: 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

90385: Achanzar WE, et al. Inorganic arsenite-induced malignant transformation of human prostate epithelial cells. J. Natl. Cancer Inst. 94: 1888-1891, 2002. PubMed: 12488483

186709: Mukta M Webber, personal communication