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Immortalized-Mouse-Distal-Caput-Epididymal-Epithelial-Cell-Line-(DC2)

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Immortalized-Mouse-Distal-Caput-Epididymal-Epithelial-Cell-Line-(DC2)

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Immortalized-Mouse-Distal-Caput-Epididymal-Epithelial-Cell-Line-(DC2)

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	Print Version
BioSafety Level	II
Organism	10-12 weeks old ts SV40 transgenic mouse
Source Organ	caput epididymis
Growth Properties	Adherent
Morphology	polygonal
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	Androgen receptor (AR), PEB-like protein, E-RABP, mEP17, cytokeratin
Applications	For Research Use Only
Immortalization Method	Isolated from a transgenice mouse carrying a temperature-sensitive simian virus 40 large T tumor antigen
Description	The epididymal epithelium plays an important role in male fertility by supporting sperm motility, maturation and viability. Owing to the changing microenvironment created by different sections of the epididymis, the spermatozoa matures as they passage through the convoluted tubule. DC2 is a conditionally immortalized mouse epididymal epithelial cell line isolated from the distal caput of the mouse harbouring a temperature-sensitive simian virus 40 large T antigen. The functional expression of the SV40 large T antigen is induced by culturing the cells in vitro at permissive temperature (33°C). At a non-permissive temperature (37°C-39°C), the cells cease to proliferate. DC2 expresses androgen receptor as well as markers of the murine epididymal epithelium, PEB-like protein (i.e. phosphatidylethanolamine binding protein), E-RABP (i.e. epididymal retinoic acid-binding protein), and EP17 (i.e. epididymal protein of 17 kd). Together with the Immortalized Mouse Proximal Caput Epididymal Epithelial Cell Line (PC1), these cells are useful in studying the regulation of tissue-specific gene expression and epididymal specific proteins in the epididymis.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Iscove Modified Dulbecco Medium (IMDM). To make the completed growth medium, add the following components to the base medium: 10% fetal bovine serum (TM999) and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 33.0°C.
Preservation	1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.
Quality Control	1) Immunocytochemistry, western blot and RT-PCR were used to confirm expression of androgen receptor, cytokeratin, mE-RABP, PEB-like protein, and mEP17; 2) RT-PCR was used to assess the T-antigen transgene expression in the immortalized cell line
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. 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 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. 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. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 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 application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.