CRL-11610 RWPE-2 人前列腺正常細胞

ATCC® Number: CRL-11610™ Price: $323.00

Designations: RWPE-2

Depositors: Michigan State University, National Cancer Institute

Biosafety Level: 2 [Cells contain Human Papilloma viral （HPV） sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens （human）

Morphology: epithelial

CRL-11610 RWPE-2 人前列腺正常細胞

Source: Organ: prostate

Disease: normal

Cell Type: epithelial

Cellular Products: cytokeratin 18 [46793]

cytokeratin 8 [46793]

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Receptors: androgen receptor, expressed （ [46793] upregulated upon exposure to androgen）

Tumorigenic: Yes

Antigen Expression: kallikrein 3, KLK3 （prostate specific antigen, PSA）; Homo sapiens, expressed （upon exposure to androgen） （ [46793] 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: At passage 14, a majority of the cells were in the near diploid range （48-54）, with the main population being 51, XY. [46795]

Isoenzymes: AK-1, 1

ES-D, 2

G6PD, B

GLO-I, 1-2

Me-2, 0

PGM1, 2

PGM3, 1

Gender: male

Ethnicity: Caucasian, White

Comments: Tumor Suppressor Gene（s）:pRB + [PubMed: 9214605]

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） [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）. RWPE-1 cells neither grow in agar nor form tumors when injected into nude mice with or without Matrigel. However, RWPE-2 cells form small colonies in soft agar and form tumors when injected into nude mice [PubMed: 9214605]. Further, 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-nitrsourea （MNU）. See the WPE1-NA22 （ATCC CRL-2849）, WPE1-NB14 （ATCC CRL-2850）, WPE1-NB11 （ATCC CRL-2851） and WPE1-NB26 （ATCC CRL-2852） cell ines.

The depositor reports that the RWPE-1 cell line （ATCC CRL-11609） was screened, and found negative for, Hepatitis B virus, Hepatitis C virus and Human immunodeficiency virus.

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

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 - 0.53mM 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 approximay 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 of 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.

Subc*tion Ratio: A subc*tion ratio of 1:3 to 1:5 is recommended

Medium Renewal: Every 2 days

Preservation: Freeze medium: Complete growth medium supplemented with 10% （v/v） DMSO

Storage temperature: liquid nitrogen vapor phase

Related Products: parental cell line:ATCC CRL-11609

purified DNA:ATCC CRL-11610D

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

derivative:ATCC CRL-2853

derived from same individual:ATCC CRL-2854

References: 46792: Webber MM, Rhim JS. Immortalized and malignant human prostatic cell lines. US Patent 5,824,488 dated Oct 20 1998

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

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

CRL-11610 RWPE-2