The attached protocol was used for growing RWPE-1 cells per ATCC instructions.

Similar documents
Human Keratinocyte Manual

Product Size Catalog Number

Thawing. 10,000-15,000 cells per cm² 10,000-15,000 cells per cm² C ,000-15,000 cells per cm². Product Size Catalog Number

Follicle Dermal Papilla Cells

Follicle Dermal Papilla Cell

To place an order, please visit lifelinecelltech.com or call customer service at

Human Mammary Luminal Epithelial Cells. Manual

Product Size Catalog Number. 500,000 proliferating cells. cryopreserved cells that have been thawed and cultured for three days at PromoCell.

RODENT Hepatocytes Care Manual

Protocol for Thawing Cryopreserved Hepatocytes

Human ipsc-derived Ventricular Cardiomyocytes. Protocol version 3.1

Endothelial Cells (Large Vessels)

Human Umbilical Vein Endothelial Cell Manual

Endothelial Cells (Microvascular)

Mouse Hepatic Progenitor Organoid Culture: Supplementary Protocols

Data Sheet IL-2-Luciferase Reporter (Luc) - Jurkat Cell Line Catalog # 60481

Data Sheet PD-1 / NFAT Reporter - Jurkat Cell Line Catalog #: 60535

Midi Plant Genomic DNA Purification Kit

PROTOCOL: OPTIMIZATION OF LENTIVIRAL TRANSDUCTION USING SPINFECTION

E.Z.N.A. SQ Blood DNA Kit II. Table of Contents

Data Sheet PD-1 / NFAT Reporter - Jurkat Cell Line Catalog #: 60535

Human Dermal Microvascular Endothelial Cells. Protocol version 1.0

L6 GLUT4myc Cell Growth Protocol

NINDS Repository Human Induced Pluripotent Stem Cell (ipsc) Handling Protocols (Matrigel and mtesr Media)

Transfection of Sf9 cells with recombinant Bacmid DNA

STAT3 (py705)/ Pan STAT3 (Human/Mouse/Rat) ELISA Kit

Notch Signaling Pathway Notch CSL Reporter HEK293 Cell line Catalog #: 60652

Large Scale Infection for Pooled Screens of shrna libraries

Product Use HPSC-CC are for research use only. It is not approved for human or animal use, or for application in in vitro diagnostic procedures.

Cryopreserved Human Hepatocytes (Cat # HP-F)

Human Induced Plutipotent Stem Cell (ipsc) Handling Protocols: Matrigel and mtesr/e8 Media

WiCell Neural Stem Cell Protocols

STAT3 (py705) (Human/Mouse/Rat) ELISA Kit

TECHNICAL BULLETIN. Catalog Number RAB0447 Storage Temperature 20 C

Extended Mammosphere Culture of Human Breast Cancer Cells

Mouse primary keratinocytes preparation

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.

Trident Membrane Protein Extraction Kit

nachr α 4 β 2 CHO Cell Line

Human Keratinocytes. Protocol version 1.0

Geneaid DNA Isolation Kit

Human Gingival Epithelial Cells. Protocol version 1.0

Cellartis DEF-CS Culture System User Manual

TECHNICAL BULLETIN. Phospho-Akt (pser 473 ) ELISA Kit for detection of human, mouse, or rat phospho-akt (pser 473 ) in cell and tissue lysates

Pluricyte Cardiomyocytes

Phosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for use with the PrimePCR Reverse Transcription Control Assay

SensoLyte pnpp Alkaline Phosphatase Assay Kit *Colorimetric*

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.

Extended Neurosphere Culture of Brain Tumor Stem Cells with the PromoCell 3D Tumorsphere Medium XF

Protocol for A-549 VIM RFP (ATCC CCL-185EMT) TGFβ1 EMT Induction and Drug Screening

Constitutive Reporter Lentiviral Vectors Expressing Fluorescent Proteins

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.

Data Sheet TIGIT / NFAT Reporter - Jurkat Cell Line Catalog #60538

Human Pluripotent Stem Cell Cardiomyocyte Differentiation Kit (PSCCDK) Introduction Kit Components Cat. # # of vials Reagent Quantity Storage

Influenza A H1N1 HA ELISA Pair Set

Macrophage Generation Media DXF

Mitochondrial DNA Isolation Kit

Immature organoids appear after hours.

STAT1 (ps727) (Human/Mouse) ELISA Kit

FOCUS SubCell. For the Enrichment of Subcellular Fractions. (Cat. # ) think proteins! think G-Biosciences

Nuclear Extraction Kit

SensoLyte 520 Cathepsin K Assay Kit *Fluorimetric*

Recombinant Trypsin, Animal Origin Free

Data Sheet. NFAT Reporter (Luc) Jurkat Cell line Catalog #: 60621

FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE.

Data Sheet. PCSK9[Biotinylated]-LDLR Binding Assay Kit Catalog # 72002

Human Immunodeficiency Virus type 1 (HIV-1) gp120 / Glycoprotein 120 ELISA Pair Set

SOP History Number Date Reason for Change v1 10/10/2014 Original V2 10/10/2016 Update V3 10/10/2018 Update

CHO α 1 β 2 γ 2 GABAA Cell Line

Protocol. G-LISA Rac1 Activation Assay Biochem Kit : 24 Assays (Absorbance Based) Cat. # BK128-S. cytoskeleton.com. Cytoskeleton, Inc.

For research or further manufacturing use only. Not for injection or diagnostic procedures.

Influenza B Hemagglutinin / HA ELISA Pair Set

Human Umbilical Cord Blood CD34 + /133+ Progenitor Cell Care Manual

20X Buffer (Tube1) 96-well microplate (12 strips) 1

EGFR (py1045)/ Pan EGFR (Human) ELISA Kit

SensoLyte Rh110 Cathepsin K Assay Kit *Fluorimetric* Revision#1.2 Last Updated: May 2017 Catalog # Kit Size

Data Sheet. CD28:B7-2[Biotinylated] Inhibitor Screening Assay Kit Catalog # Size: 96 reactions

ACTG Laboratory Technologist Committee Revised Version 2.0 ACTG Lab Man HIV Syncytium-Inducing (MT-2) assay 29 April 2004

Thawing MEFs (Mouse Embryonic Fibroblasts (MEFs)

Mammalian Membrane Protein Extraction Kit

Human TSH ELISA Kit. User Manual

Primary Adult Naïve CD4+ CD45RA+ Cells. Prepared by: David Randolph at University of Alabama, Birmingham

Mitochondrial Trifunctional Protein (TFP) Protein Quantity Microplate Assay Kit

Human CD4+T Cell Care Manual

Instructions. Fuse-It-mRNA easy. Shipping and Storage. Overview. Kit Contents. Specifications. Note: Important Guidelines

91154 PRIME-XV T Cell CDM Liquid 1 L Additional package sizes are available at request

Data Sheet. PCSK9[Biotinylated]-LDLR Binding Assay Kit Catalog # 72002

Human Urokinase / PLAU / UPA ELISA Pair Set

A protocol for enhancement of the AAV-mediated expression of transgenes

Corning Cryopreserved HepatoCells Instructions for Use

CytoSelect Tumor- Endothelium Adhesion Assay

Biological Consulting Services

Gladstone Institutes, University of California (UCSF), San Francisco, USA

HIV-1 Virus-like Particle Budding Assay Nathan H Vande Burgt, Luis J Cocka * and Paul Bates

1.1. Gelatinizing Plates Prepare plates by covering surface with 0.1% Gelatin solution:

Exo-spin Exosome Purification Kit For cell culture media/urine/saliva and other low-protein biological fluids

Influenza A H7N9 (A/Anhui/1/2013) Hemagglutinin / HA ELISA Pair Set

ACTG Laboratory Technologist Committee Revised Version 2.0 ACTG Lab Man HIV Qualitative PBMC Micrococulture 1 June 2004

ab Adipogenesis Assay Kit (Cell-Based)

Human Immunodeficiency Virus type 1 (HIV-1) p24 / Capsid Protein p24 ELISA Pair Set

Transcription:

RWPE-1 (prostate epithelial transformed by HPV) From: Duke/UNC/UT/EBI ENCODE group Date: 7/7/11 ATCC: Catalog #CRL-11609 The attached protocol was used for growing RWPE-1 cells per ATCC instructions.

Cell Biology ATCC Number: CRL-11609 Order this Item Price: $279.00 Designations: RWPE-1 Depositors: Michigan State University, National Cancer Institute Biosafety Level: 2 [Cells contain Human Papilloma viral (HPV) sequences ] Shipped: Medium & Serum: Growth Properties: Organism: Morphology: Source: Cellular Products: Permits/Forms: Receptors: Tumorigenic: Antigen Expression: DNA Profile (STR): Cytogenetic Analysis: frozen See Propagation adherent Homo sapiens (human) epithelial Organ: prostate Disease: normal Cell Type: epithelial cytokeratin 18 [46793] cytokeratin 8 [46793] 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. androgen receptor, expressed ( [46793] upregulated upon exposure to androgen) No kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens, expressed (upon exposure to androgen) ( [46793] upon exposure to androgen) 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 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] Related Links NCBI Entrez Search Cell Micrograph Make a Deposit Frequently Asked Questions Material Transfer Agreement Technical Support Related Cell Culture Products Login Required Product Information Sheet BioProducts Cell, microbial and molecular genomics products for the life sciences BioServices Bio-materials management; basic repository to complex partnership-level services

Isoenzymes: Age: Gender: Ethnicity: Comments: AK-1, 1 ES-D, 2 G6PD, B GLO-I, 1-2 Me-2, 0 PGM1, 2 PGM3, 1 54 years adult male Caucasian, White Tumor Supressor Gene(s): p53 + [PubMed: 9214605] prb + [PubMed: 9214605] Epithelial cells derived from the peripheral zone of a histologically normal adult human prostate were transfected with a single copy of the human papilloma virus 18 (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. BioStandards Biological Reference Material and Consensus Standards for the life science community

Propagation: Subculturing: Preservation: 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 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 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 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 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 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 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 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 hormonerelated 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

Return to Top