The renal proximal tubule is part of the nephron duct system in the kidney, playing important roles in functions like absorption and secretion of molecules in the renal system. The Immortalized Mouse Renal Proximal Tubule Cells is a conditionally immortalized mouse renal proximal tubule cells isolated from the mouse (C57BL/6J x Immortomouse, designated as CBA;B10-Tg(H2Kb-tsA58)6Kio/Crl) harboring thermolabile mutation (tsA58) of the simian virus 40 large T antigen. The functional expression of the SV40 large T antigen is induced by culturing the cells in vitro in medium containing IFN gamma at a temperature permissive (33°C). At a non-permissive temperature (37°C-39°C), the cells cease to proliferate. These cells retain competent transepithelial electrolyte transport and short-circuit current activities. When paired up with angiotensin receptor (Ang II)-deficient cell lines AT1A -/- (Cat. No. T0626), AT1B -/- (Cat. No. T0627), AT1A -/- AT1B -/- (Cat. No. T0628) and AT2 -/- (Cat. No. T0625), these cell lines represent valuable tools to study fluid and electrolyte transportation in the proximal tubule region.
Cell Type
Immortalized Cells
Disclaimer
1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item. 2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 3. 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 mug, Cat.# C207, $450.00) or cell lysate (100 mug, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 4. 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). 5. 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. 6. 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. 7. 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."
Growth Properties
Adherent
Quantity
5x105 cells / 1.0 ml
Tissue
Kidney
Morphology
Cobble-stone
Species
Mouse (M. musculus)
Propagation
Use of Millicell cell culture inserts are required for cell adhesion and growth. Coat these inserts with Applied Cell Extracellular Matrix (G422) before use.The base medium 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 final concentration of 5%, aldosterone to a final concentration of 10 nM, L-ascorbic acid 2-phosphate to a final concentration of 50 µM, dexamethasone to a final concentration of 4 µg/mL, epidermal growth factor (Z200025) to a final concentration of 10 ng/mL, insulin (Z101065) to a final concentration of 5 µg/ml, sodium selenite to a final concentration of 20 nM, transferrin to a final concentration of 5 µg/mL, L-3,3,5-triiodothyronine (T3) to a final concentration of 1 nM, recombinant mouse IFNG (Z200085) to a final concentration of 10 U/mL, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: 33.0°C.* Do not use heat-inactivated FBS for cell culture unless specified otherwise.To differentiate the cells, grow the cells at 37°C-39°C in the absence of IFNG.
Quality Control
(1) RT-PCR was performed to confirm AT receptor genotype. (2) Electrical conductivity was measured to compare immortalized cells relative to non-immortalized counterparts. (3) Immunostaining was performed to determine localization of proteins associated with epithelial cells and assess morphology. (4) Changes in short circuit current were quantified to compare immortalized cell cotransporters to non-immortalized ones. (5) Western blotting was performed to detect the presence of AT receptor proteins. (6) Live cell microscopy of AngII-mediated beta-arrestin 2 translocation was performed to measure receptor functionality.
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