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. Stromal smooth muscle cells established in primary culture after radical prostatectomy from a 62-year-old patient were subjected to retroviral infection with human telomerase catalytic subunit (hTERT), thus creating Immortalized Human Prostate Stromal Cells (PM151T) - hTERT. These cells were able to proliferate for at least 190 days, express genes associated with smooth muscle function (progesterone receptor, estrogen receptor-β, androgen receptor, smooth muscle-myosin heavy chain, calponin, SM22, smooth muscle α-actin and myocardin) and exhibit up-regulation for some of these markers as well as contractile behavior upon induction of differentiation. 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: II
Species: Human
Source Organ: Prostate
Growth Properties: Adherent
Morphology: Fibroblast
Population Doubling: N/A
Seeding Density: 2500 cells/cm²
Markers: Puromycin resistance, progesterone receptor, estrogen receptor-β, androgen receptor, smooth muscle-myosin heavy chain, calponin, SM22, smooth muscle α-actin, myocardin
Donor Age: 62 years old
Donor Gender: Male
Donor Ethnicity: N/A
Propagation: The base medium for this cell line is MCDB 131 medium (Sigma). To make the complete growth medium, add the following components to the base medium: decalcified horse serum (Sigma) to a final concentration of 15%, pH 7.2 HEPES to a final concentration of 10mM, MEM Non-essential Amino Acid Solution to a final concentration of 1X, insulin (Sigma) to a final concentration of 5 µg/mL, transferrin (Sigma) to a final concentration of 10 µg/mL, estradiol to a final concentration of 10 nM, dexamethasone (Sigma) to a final concentration of 100 nM, sodium selenite (Sigma) to a final concentration of 5 ng/mL, basic FGF (Sigma) to a final concentration of 1 ng/mL and EGF (Sigma) to a final concentration of 0.1 ng/mL. Change media every 2-3 days. Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
* Do not use heat-inactivated serum for cell culture unless specified otherwise.
Quality Control: 1) Western blot; 2) qRT-PCR; 3) Transforming growth factor-β/endothelin A treatment
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C