Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Organism	Homo sapiens, human
Tissue	prostate/stroma
Cell Type	Epithelial, fibroblast, Myofibroblast
Product Format	frozen
Morphology	myofibroblast
Culture Properties	adherent
Biosafety Level	2 [Cells contain SV40 viral DNA sequence] Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	normal
Age	54 years
Gender	male
Ethnicity	Caucasian
Applications	Because of their derivation from the same peripheral zone of the prostate, the WPMY-1 cell line is especially useful for studies on paracrine and stromal : epithelial interactions.
Storage Conditions	liquid nitrogen vapor phase

Karyotype	At passage 66, a majority of the cells were in the 58-68 range; X, -Y. Ref
Derivation	The myofibroblast stromal cell line, WPMY-1, was derived from stromal cells from the same peripheral zone of the histologically normal adult prostate, as that used for RWPE-1 cells (ATCC CRL-11609). Stromal cells were immortalized with SV40-large-T antigen gene, using a pRSTV plasmid construct. WPMY-1 stromal cells belong to a family of cell lines derived from the same prostate as the epithelial RWPE-1 cells and all of its epithelial derivatives.
Clinical Data	54 years Caucasian, White male
Antigen Expression	kallikrein 3, KLK3 (prostate specific antigen, PSA); Homo sapiens Ref
Receptor Expression	androgen receptor, expressed Ref
Genes Expressed	fibronectin Ref smooth muscle alpha-actin vimentin Ref
Cellular Products	fibronectin Ref smooth muscle alpha-actin vimentin Ref
Tumorigenic	No
Effects	No, into nude mice (at Passage 22 cells did not form tumors when injected subcutaneously)(Mukta M Webber, personal communication) Yes, the cells form colonies in soft agar (colony forming efficiency (CFE) of 0.7%)(Mukta M Webber, personal communication)
Comments	The depositor reports that the RWPE-1 cell line (ATCC CRL-11609), derived from the same prostate, was screened for Hepatitis B and C, and human immunodeficiency viruses, and was found to be negative.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated Dulbecco''s Modified Eagle''s Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 5%.
Subculturing	Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Note: Subculture cells before or upon reaching confluence. Do not allow cells to become super-confluent. 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 of 0.025% Trypsin - 0.26 mM EDTA solution (1:1 dilution of 0.05% Trypsin - 0.53 mM EDTA in D-PBS) to the flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping, do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of 0.1% Soybean Trypsin Inhibitor (or 2% fetal bovine serum in D-PBS) and aspirate cells by gently pipetting. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum between 1 x 104 to 2 x 104 viable cells/cm2 is recommended. Incubate cultures at 37°C. We recommend that you maintain cultures at a cell concentraton between 4 x 104and 8 x 104 cells/cm2. Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: Every 48 hours
Cryopreservation	Freeze medium: Complete growth medium supplemented with an additional 15% fetal bovine serum and 10% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C Growth Conditions: Subculture cells before or upon reaching confluence. Do not allow cells to become super-confluent.