Description: Prostate cancer research has been hindered by the fact that the cell types involved in the process of malignant transformation have proven difficult to develop into continuously growing cultures. Epithelial cells established in primary culture after radical prostatectomy from a 66-year-old patient were subjected to retroviral infection with human telomerase catalytic subunit (hTERT), thus creating the Immortalized Human Prostate Epithelial Cells (EP156T) - hTERT line. These cells underwent up to 200 population doublings, express genes consistent with a prostate basal epithelial phenotype (cytokeratin 5/6/7/8/14/18, p63, estrogen receptor-β and androgen receptor) and, like primary cells, differentiate as spheroids with a secretory structure when grown in three-dimensional culture. The generation of this cell line by way of a method that minimizes complex alterations results in a faithful in vitro model for the prostate, offering a tool for the study of not only normal development and physiology, but also the steps leading to malignancy.Immortalization Method: Immortalized with retrovirus containing hTERT.BioSafety Level: IISpecies: HumanSource Organ: ProstateGrowth Properties: AdherentMorphology: EpithelialPopulation Doubling: N/ASeeding Density: 2500 cells/cm10²Markers: Puromycin resistance, cytokeratin 5/6/7/8/14/18, p63, estrogen receptor-β, androgen receptorDonor Age: 66 years oldDonor Gender: MaleDonor Ethnicity: N/APropagation: The base medium for this cell line is MCDB 153 medium (Sigma). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 1%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, MEM Non-essential Amino Acid Solution to a final concentration of 1X, bovine pituitary extract (Gibco) to a final concentration of 50 µg/mL, insulin (Sigma) to a final concentration of 5 ug/mL, transferrin (Sigma) to a final concentration of 5 ug/mL, EGF (Sigma) to a final concentration of 5 ng/mL, hydrocortisone to a final concentration of 200 nM, triiodothyronine (Sigma) to a final concentration of 10 nM, testosterone (Sigma) to a final concentration of 10 nM and sodium selenite (Sigma) to a final concentration of 5 ng/mL.
Change media every 2-3 days.
Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
* Do not use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: 1) Western blot; 2) qRT-PCR; 3) Three-dimensional cell cultureShipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C