Immortalized Human Prostate Stromal Cells (PM151T) - hTERT

Mfr.No.	T0727
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. 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.?br />These cells were able to proliferate for at least 190 days, express genes associated with smooth muscle function (progesterone receptor, estrogen receptor-?, androgen receptor, smooth muscle-myosin heavy chain, calponin, SM22, smooth muscle ?-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. Characterization by Western blot, qRT-PCR, and transforming growth factor-?/endothelin A treatment.
Cell Type	Immortalized Cells
Biosafety Level	2
Depositor	Yeda Research and Development Co. Ltd.
Disclaimer	1. All test parameters provided in the CoA are conducted using abm's standardized culture system and The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic or any other non-RUO application(s). 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period".
Donor History	Male, 62
Expression Region	Progesterone receptor, estrogen receptor-?, androgen receptor, smooth muscle-myosin heavy chain, calponin, SM22, smooth muscle ?-actin, myocardin, puromycin-resistance
Expression Level	NBCe1-A, NHE3
Growth Properties	Adherent, fibroblast
Growth Conditions	Use of PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. MCDB 131 medium (Sigma) + 15% decalcified horse serum (Sigma) + 10 mM pH 7.2 HEPES + 1X MEM Non-essential Amino Acid Solution + 5 ?/ml insulin (TM053) + 10 ?/ml transferrin (Sigma) + 10 nM estradiol + 100 nM dexamethasone (Sigma) + 5 ng/ml sodium selenite (Sigma) + 1 ng/ml basic FGF (Z101456) + 0.1 ng/ml EGF (Z100139) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?
Quantity	5x105 cells / 1.0 ml
Tissue	Prostate
Notes	1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use.To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Human (H. sapiens)
Population Doubling	24 - 48 hours
Species	Human (H. sapiens)
Propagation	Cells are grown on Millicell Cell Culture Insert, 12 mm, hydrophilic PTFE, 0.4 mum (Millipore) that are lightlycoated with a 20% dispersion of rat-tail collagen (in 60% ethanol) plus 1 mug/mL laminin.The basemedium for this cell line is Prigrow IV medium available at?abm, Cat.No.?TM004. To make the complete growth medium,add the following components to the base medium: fetal bovine serum (TM999)* to a finalconcentration of 5%, HEPES (Sigma) to a final concentration of 15 mM, sodium selenite (Sigma) to a finalconcentration of 20 nM, ascorbic acid-2-phosphate (Sigma) to a final concentration of 50 mM, transferrin (Sigma)to a final concentration of 5 mug/mL, Insulin Solution (TM053) to a final concentration of 5 mug/mL, mouseEGF (Sigma) to a final concentration of 10 ng/mL, dexamethasone to a final concentration of 4 mug/mL, T3 (Sigma)to a final concentration of 1 nM, and Penicillin/Streptomycin Solution (G255) to a finalconcentration of 1%.Change media every 2-3 days by first removing it from the bottom of thefilter and then from the top. Add fresh medium first to the top and then to the bottom.Carbondioxide (CO2): 5%, Temperature: 33.0°C. To differentiate cells, increasetemperature to 38.0°C.* Do not use heat-inactivated FBS for cell culture unless specifiedotherwise.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control	1) Western blot

Applications Range	Research Use Only.

Shipping	Ship with dry ice.
Product Format	Frozen
Product Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol	Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10900678	T0727	1x10^6 cells / 1.0 ml	-		Add to Quote

Immortalized Human Prostate Stromal Cells (PM151T) - hTERT

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