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運輸方式：凍存運輸
生長狀態(tài)：貼壁生長
年限：54
ATCC Number：CRL-2887?
器官來源：其他
細胞類型：上皮細胞
是否是腫瘤細胞：0
物種來源：人
數(shù)量：大量
細胞形態(tài)：上皮樣
規(guī)格：0.2ml Designations: WPE-stem
Depositors: ?MM Webber, EJ Tokar
Biosafety Level:2 [cells contain human HPV-18 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:loosely adherent
Organism: Homo sapiens
Morphology:epithelial


Source: Organ: prostate, normal, peripheral zone
Cell Type: epithelial
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: Isolation date: 2004 [PubMed: 16351690]
Applications:WPE-stem cells are loosely adherent and exhibit features characteristic of stem/progenitor cells present in the embryonic urogenital sinus and in adult prostatic epithelium, including p63 and ABCG2. The cells show high expression of cytokeratin 5 and 14 and MMP-2 but low expression of cytokeratin 18. They are androgen-independent for growth and survival.
Receptors:androgen receptor expressed, slightly upregulated upon exposure to androgen [PubMed: 16351690] [92794 ]
Tumorigenic:No
Antigen Expression:Weak expression of prostate specific antigen (PSA) and androgen receptor (AR) upon exposure to androgen. [PubMed: 16351690].
DNA Profile (STR):Amelogenin: X,Y
CSF1PO: 13
D13S317: 8,14
D16S539: 9,11
D5S818: 12,15
D7S820: 10,11
THO1: 8
TPOX: 8,11
vWA: 14,18
Age: 54
Gender: male
Ethnicity: Caucasian
Comments:WPE-stem cells were derived from the RWPE-1 cell line (ATCC CRL-11609 ) after two consecutive cycles of single cell cloning. To establish the RWPE-1 cell line, 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. WPE-stem cells are loosely adherent and exhibit features characteristic of stem/progenitor cells present in the embryonic urogenital sinus and in adult prostatic epithelium, including p63 and ABCG2. The cells show high expression of cytokeratin 5 and 14 and MMP-2 but low expression of cytokeratin 18. They are androgen-independent for growth and survival. Cells grow in soft agar with a cloning efficiency of 0.9%. Cells are not tumorigenic in nude mice even at 6 months after injection. The WPE-int cell line (ATCC CRL-2888 ), which has an intermediate phenotype on the path to luminal cell differentiation, was also derived from RWPE-1 cells after single cell cloning. [PubMed: 16351690]

WPE-stem cells were screened for CMV, HBV, HCV, HTLV 1, HTLV 2, HIV 1, HIV 2, JCV, and MoMuLV DNA sequences. Cells were also tested for 25 species of mycoplasma and Acholeplasma laidlawii . Cells tested negative for all of the above. (personal communication from depositor).

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°C
Growth Conditions: Note: Culture flasks should be pre-coated with a mixture of Mouse Collagen Type IV and Human Fibronectin (Sigma Cat No. F-0895) 2.5 microgram each/sq. cm.

Subculturing: Protocol:

Volumes used in this protocol are for 75 sq.cm flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Subculture cells before they reach confluence. Do not allow cells to become confluent.
Note: Culture flasks should be pre-coated with a mixture of Mouse Collagen Type IV and Human Fibronectin (Sigma Cat. No. F-0895) 2.5?g each/ sq.cm.
1. Collect spent medium containing viable, floating, single cells, and prostaspheres.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).
3. 3.0 to 4.0 ml of 0.05% Trypsin - 0.53 mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37°C incubator for 3 to 5 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 8 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, add the spent medium containing floating cells, and aspirate cells by gently pipetting.
5. Transfer cell suspension to centrifuge tube and spin at approximately 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 pre-coated culture flasks. An inoculum of 2 X 10(3) to 4 X 10(3) viable cells/sq. cm is recommended.
7. Place culture vessels in incubators at 37°C. 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.
   Interval: Subculture cells before they reach confluence. Do not allow cells to become confluent. Subculture when cell concentration is between 1 X 10(5) and 1.5 X 10(5) cells/sq. cm.
   Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended
   Medium Renewal: Every 48 hours

Preservation: Freeze medium: Complete growth medium described above supplemented with 15% fetal bovine serum and 10% (v/v) DMSO. Cell culture tested DMSO is available as ATCC ? Catalog No. 4-X.
Storage temperature: liquid nitrogen vapor phase
Doubling Time: about 24 hours
Related Products:recommended serum: ATCC 30-2020
phosphate-buffered saline: ATCC 30-2200
Cell culture tested DMSO: ATCC 4-X
Erythrosin B vital stain solution: ATCC 30-2404
parental cell line: ATCC CRL-11609
derived from the 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-2851
derived from same individual:ATCC CRL-2852
derived from same individual:ATCC CRL-2853
derived from same individual:ATCC CRL-2854
derived from same individual:ATCC CRL-2888
derived from same individual:ATCC CRL-2889
derived from same individual:ATCC CRL-2890
derived from an unrelated individual: ATCC CRL-11611
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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
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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
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
92794: Tokar EJ, et al. Stem/progenitor and intermediate cell types and the origin of human prostate cancer. Differentiation 73: 463-473, 2005. PubMed: 16351690
92795: Achanzar WE, et al. Human prostate cell lines mimic heterogeneity of cadhedrin expression in human prostate cancer. Urol. Oncol. 4: 15-25, 2004.
16172794: Bramblia 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.