Immortalized Mouse Renal Proximal Tubule Cells-Conditionally

T06245x105 cells / 1.0 ml



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 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.

SpeciesMouse (M. musculus)
Species descriptionC57BL/6J x Immortomouse
Tissue/Organ/Organ SystemKidney
Growth PropertiesAdherent
Cell MorphologyCobble-stone
Immortalization MethodIsolated from transgenic mouse (C57BL/6J x Immortomouse) carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)

For Research Use Only

Unit quantity5x105 cells / 1.0 ml
Cell TypeImmortalized Cells
Propagation Requirements

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.


(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.


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 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.

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."

DepositorCase Western Reserve University

I want to make sure these cells express my gene of interest before I decide to buy the cell line. Can you provide a sample so this can be tested?
We do not carry out downstream characterization or gene expression profiling of our cell lines. To facilitate your preliminary experiments we can provide an RNA extraction (0.5ug total RNA) or cell lysate (100ug/100ul provided in 62.5mM Tris‐HCl, 2% SDS, 10% Glycerol, 50mM DTT, 0.01% w/v Bromophenol Blue) for any of our immortalized cell lines for a small fee. Please inquire directly for more information. The lead time will be around 2 weeks from the time of placing an order (if the item is in stock).
How often do I need to change the media?
The media should be changed every 2-3 days.
Why do these cells need bio safety level II?
In order to be more cautious, we follow the CDC-NIH recommendations that all mammalian sourced products should be handled at the Biological Safety Level 2 to minimize exposure of potentially infectious products. This information can be found in 'Biosafety in Microbiological and Biomedical Laboratories' (1999). Your institution's Safety Officer or Technical Services will be able to make the call as to whether BioSafety Level I is possible with these cells at your site if required.
Do you sell ECM coated T75 flasks?
Yes we can provide a coating service. Please inquire with [email protected]
What can I coat a larger dish to subculture?
We also offer applied extracellular matrix (collagen type I) in liquid form, for the coating of larger flasks and other required plasticware:
How long can I store frozen vials for?
Cells that are properly frozen using an effective cryoprotective agent can be stored in liquid nitrogen indefinitely without affecting their recovery.
Should the cap of the flask be changed before starting the cell culturing step?
No, there is no need in sterile biosafety cabinets unless it has contacted any non-sterile condition (e.g. touching the contaminated tip, etc.).
What is the recommended storage temperature?
In general, if you received: Live cells: acclimatize for 3-4 hrs at at the recommended conditions stated for the cell line under the propagation section, and then change media afterwards. Frozen cells: Immediately place cells in liquid nitrogen; -180C.
How is cell density crucial for drug selection?
If antibiotic selection is applicable to the target cells, we suggest getting rid of all the background cells so that the cell density is kept lower (even 20-30%). However, once the clones are selected by clonal dilution, we don't need the drug to still be present. If needed, the cell density should be towards the higher end since cells are already selected. Any primary cells still present will be depleted as a result of senescence and the cell population that remains will be resistant to the specific antibiotic.
My cells are not detaching, what method do you recommend to trypsinize the cells?
1. Incubate the coated plate containing trypsin solution at recommended temperature indicated in the propagation section for 3-5 min till the cells round up, monitoring from time to time under microscope. 2. Diluting G422 (1:1) with PBS and coating for lesser time. Sometimes the collagen content in G422 is higher and thus make stronger bonding with cells. 3. You can try reducing the incubation time as well for coating the plate to make a thinner layer.
Why is it important to determine the optimal seeding density?
The seeding density we recommend is for when cells are plated to a new vessel. The optimal seeding density should allow cells to attach to the surface and have room to proliferate. If you seed too little, cells may not attach well to the surface (for adherent cells). Seeding density is important as many cells (adherent or suspension cells) need to be in close proximity for better growth. Cell-cell interactions allow cells to communicate with each other in response to changes in their microenvironment. This ability to send and receive signals is essential for the survival of the cell. In other cases, if the seeding density is too low, cells may attach but a retardation in cell growth is observed. If you seed too high, the cells will attach but there is insufficient room for further proliferation and they will stop replicating.
What is the construct used to make the immortalization?
The construct used contain H-2Kb 5' promoter sequences fused to the early region of the SV40 mutant tsA58. The thermolabile mutation (tsA58) simian virus 40 large T antigen makes this cell line conditionally immortalized under permissive temperature of 33°C. At a non-permissive temperature (37°C-39°C), the cells cease to proliferate.

  • Woost, PG et al. "Strategy for the development of a matched set of transport-competent, angiotensin receptor-deficient proximal tubule cell lines." In Vitro Cell Dev Biol Anim 42(7):189-200 (2006). PubMed: 16948500 .
  • Li, XC et al. "Mechanisms of AT1a receptor-mediated uptake of angiotensin II by proximal tubule cells: a novel role of the multiligand endocytic receptor megalin" Am J Physiol Renal Physiol. 307(2):F222-33 (2014). PubMed: 24740791.
  • Li, XC et al. "Role of caveolin 1 in AT1a receptor-mediated uptake of angiotensin II in the proximal tubule of the kidney" Am J Physiol Renal Physiol. 307(8):F949-61 (2014). PubMed: 25164083 .
  • Li, XC et al. "Novel signaling mechanisms of intracellular angiotensin II-induced NHE3 expression and activation in mouse proximal tubule cells" Am J Physiol Renal Physiol. 303(12):F1617-28 (2012). PubMed: 23034941 .
  • Li, XC et al. "AT1 receptor-mediated uptake of angiotensin II and NHE-3 expression in proximal tubule cells through a microtubule-dependent endocytic pathway" Am J Physiol Renal Physiol. 297(5):F1342–F1352 (2009). PubMed: PMC2781332.
  • Noble, JP et al. "Direct derivation of conditionally immortal cell lines from an H-2Kb-tsA58 transgenic mouse" Proc Natl Acad Sci USA 88(12):5096-5100 (1991). PubMed: 1711218.
  • Woost, PG et al. "Development of an AT2-deficient proximal tubule cell line for transport studies" In Vitro Cell Dev Biol Anim. 43(10):352-360 (2007). PubMed: 17963016.