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Immortalized-Mouse-Cortical-Collecting-Duct-Cell-Line-(mCCD(cl1))

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Immortalized-Mouse-Cortical-Collecting-Duct-Cell-Line-(mCCD(cl1))

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Immortalized-Mouse-Cortical-Collecting-Duct-Cell-Line-(mCCD(cl1))

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	Print Version
Biosesafety Level	II
Organism	Mus musculus
Source Organ	Kidney
Growth Properties	Adherent
Recommended Seeding Density	In early passages, the cells do not form domes and grow more slowly. Split low passage cells at 1:6 ratio. At higher passages (above p15), the cells grow more rapidly and are recommended to be split at 1:10 ratio.
Applications	For Research Use Only
Immortalization Method	Spontaneously immortalization
Cell Type Characterization
Description	In aldosterone-sensitive distal nephron (ASDN), mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) activation regulate ionic and water transports through the epithelial sodium channel (ENaC) and the Na+/ K+ pump. The Immortalized Mouse Cortical Collecting Duct Cell Line is a spontaneously transformed cell line that expresses significant level of functionally active MR and GR, as well as active 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) enzyme. In addition, this cell line responses to physiological concentrations (K(1/2)= 0.3 nM) of aldosterone, making it useful for binding equilibrium assays and studies on sodium transport response to various mineralocorticoid and glucocorticoid agonists.
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 Prigrow IV medium available from ABM (TM004). To make the complete growth medium, add the following components to the base medium: 2% fetal calf serum, 0.87 µM insulin, 5 µg/ml human apotransferrin, 10 ng/ml EGF, 1 nM T3, 30 nM dexamethasone and 1% Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO2: 5%; Temperature: 37.0°C.
Quality Control	1) Immunofluorescence and RT-PCR were used to confirm the expression of epithelial sodium channel (ENaC) and Na-K-ATPase; 2) RT-PCR was used to detect expression of MR, GR, and 11β-HSD2 ; 3) sodium transport response assayed through short-circuit current (electrophysiologic studies) and binding assays.
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 evalsuate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical locations. 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.