RWPE-1細(xì)胞

ATCC Number： CRL-11609?

相關(guān)**： 正常

細(xì)胞類型： 上皮細(xì)胞

生長狀態(tài)： 貼壁生長

是否是腫瘤細(xì)胞： 0

物種來源： 人

年限： 54 years *****

運(yùn)輸方式： 凍存運(yùn)輸

數(shù)量： 大量

器官來源： 前列腺

細(xì)胞形態(tài)： 上皮樣

Designations: RWPE-1

Depositors: Michigan State University, National Cancer Institute

RWPE-1細(xì)胞Biosafety Level: 2 [Cells contain Human Papilloma viral (HPV) sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens deposited as human

Morphology: epithelial

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: No

Antigen Expression: kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens, expressed (upon exposure to androgen) ( [46793] upon exposure to androgen)

RWPE-1細(xì)胞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 32, a majority of the cells were in the diploid range (45-51) with two main populations: 45, X,-Y and 51, XY. [46793]

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: Tumor Supressor Gene(s):

p53 + [PubMed: 9214605]

pRB + [PubMed: 9214605]

Epithelial cells derived from the peripheral zone of a histologically normal ***** human prostate were transfected with a single copy of the human papilloma virus 18RWPE-1細(xì)胞 (HPV-18) to establish the RWPE-1 (ATCC CRL-11609) cell line [PubMed: 9214605]. In 3-dimensional Matrigel culture, RWPE-1 cells organize into acini and secrete PSA into the lumen when exposed to androgen [PubMed: 11170142]. When injected with Matrigel or with stromal cells, into male athymic rodents, RWPE-1 cells also organize into acini [PubMed: 11304724] and produce PSA. Cells from the RWPE-1 cell line were further transformed by Ki-ras using the Kirstin murine sarcoma virus (Ki-MuSV) to establish the tumorigenic RWPE-2 cell line (ATCC CRL-11610) [PubMed: 9214605] and the RWPE2-W99 (ATCC CRL-2853) cell line. 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-nitrosourea (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 lines.

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:

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 - 0.53mM 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.

Add 6.0 to 8.0 ml of 0.1% Soybean Trypsin InhibitorRWPE-1細(xì)胞 (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 of 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 7 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 2 days

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

Storage temperature: liquid nitrogen vapor phase

Related Products: derivative:ATCC CRL-11610

purified DNA:ATCC CRL-11609D

derivative:ATCC CRL-2849

derivative:ATCC CRL-2850

derivative:ATCC CRL-2852

derivative:ATCC CRL-2851

derived from same individual: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 ***** 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. RWPE-1細(xì)胞PubMed: 9214606

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

RWPE-1細(xì)胞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