Immortalized Mouse Distal Caput Epididymal Epithelial Cell Line (DC2)

Cat. No.
T0599
Unit
1x10⁶ cells / 1.0 ml
Price
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Cat. No. T0599
Name Immortalized Mouse Distal Caput Epididymal Epithelial Cell Line (DC2)
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) (Cat. No. T0598), these cells are useful in studying the regulation of tissue-specific gene expression and epididymal specific proteins in the epididymis. This cell line may contain some fibroblast cells. Immunocytochemistry, western blot and RT-PCR were used to confirm expression of markers.


Unit 1x10⁶ cells / 1.0 ml
Cell Type Immortalized Cells
Organism Mouse (M. musculus)
Tissue Testes
Donor History 10-12 weeks old tsSV40 transgenic mouse, Caput epididymis
Growth Properties Adherent, polygonal
Seeding Density (cells/cm2) 20,000 - 30,000
Population Doubling Time (h) 70 - 80
Immortalization Method Isolated from a transgenic mouse carrying a temperature-sensitive simian virus 40 large T tumor antigen
Expression Profile

Androgen receptor (AR), PEB-like protein, E-RABP, mEP17, cytokeratin

Growth Conditions Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow V (TM015) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 33.0°C, 5% CO₂
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°C, 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.

QC 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
Application Research Use Only.
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
BioSafety II
Disclaimer
1. 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 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.

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."
Depositor Vanderbilt University
Print & Download Datasheet
  • Araki, Y., Suzuki, K., Matusik, R. J., Obinata, M., & Orgebin-Crist, M. C. (2002). Immortalized epididymal cell lines from transgenic mice overexpressing temperature-sensitive simian virus 40 large T-antigen gene. Journal of andrology, 23(6), 854–869.


    Chan, J. C., Morgan, C. P., Adrian Leu, N., Shetty, A., Cisse, Y. M., Nugent, B. M., … Bale, T. L. (2020). Reproductive tract extracellular vesicles are sufficient to transmit intergenerational stress and program neurodevelopment. Nature Communications, 11(1). doi:10.1038/s41467-020-15305-w

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