Immortalized Mouse Renal Proximal Tubule Cells-Conditionally

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 γ 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 AT_1A ^-/- (Cat. No. T0626), AT_1B ^-/- (Cat. No. T0627), AT_1A ^-/- AT_1B ^-/- (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.

Immortalization Method: Isolated from transgenic mouse (C57BL/6J x Immortomouse) carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)

BioSafety Level: II

Species: Mouse

Source Organ: Kidney

Growth Properties: Adherent

Morphology: Cobble-stone

Population Doubling: N/A

Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.

Markers: N/A

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow IV medium available at abm (TM004). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 5%, 10 nM aldosterone, 50 µM L-ascorbic acid 2-phosphate, 4 µg/mL dexamethasone, 10 ng/mL epidermal growth factor, 5 µg/mL insulin, 20nM sodium selenite (Na2SeO3), 5 µg/mL transferrin, 1 nM L-3,3’,5-triiodothyronine (T3), 10 U/mL recombinant mouse interferon gamma (IFN-γ) and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO₂): 5%, Temperature: +33.0°C. To differentiate the cells, grow the cells at +37°C-39°C in the absence of IFN-γ. * abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

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 β-arrestin 2 translocation was performed to measure receptor functionality.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C