RLE-6TN Cells

Categories: Animal cells Description: The RLE-6TN cell line is an immortalized rat alveolar type II epithelial cell line derived from adult Fischer 344 rats. RLE-6TN was established through spontaneous immortalization during attempts to introduce the SV40-T antigen gene into primary alveolar type II epithelial cells. Unlike its counterpart RLE-6T, which was positively transfected with the SV40-T antigen, RLE-6TN cells do not express the T-antigen gene. Despite this, RLE-6TN cells retain critical morphological and functional features characteristic of alveolar type II cells, including cytokeratin expression and the presence of lipid-containing lamellar inclusion bodies. RLE-6TN cells have been widely used as an in vitro model for investigating lung epithelial cell biology, alveolar function, and responses to various physiological and pathological stimuli. They are particularly relevant for studying Na-K-ATPase regulation and activity in alveolar epithelial cells. Na-K-ATPase is essential for maintaining cellular ion gradients and trans-epithelial ion transport, processes critical for alveolar fluid clearance in the lungs. In studies, thyroid hormone (T3) has been shown to stimulate Na-K-ATPase activity in RLE-6TN cells by enhancing its translocation to the plasma membrane rather than increasing its transcription, highlighting a novel, rapid regulatory mechanism. RLE-6TN cells demonstrate stable growth, with near-diploid karyotype stability, and are not tumorigenic in nude mice. They are negative for alkaline phosphatase activity but stain positive for cytokeratins 8, 18, and 19, confirming their epithelial origin. RLE-6TN cells can be maintained long-term in culture and serve as a reliable platform for mechanistic studies on alveolar epithelial repair, surfactant metabolism, and cellular responses to lung injury, toxins, and therapeutic agents. Organism: Rat Tissue: Lung Synonyms: Rat Lung Epithelial-6-T-antigen Negative Age: 56 days Gender: Male Morphology: Epithelial Growth Properties: Adherent Citation: RLE-6TN (Cytion catalog number 305350) Biosafety Level: 1 Ncbi_ Taxid: 10116.0 Cellosaurus Accession: CVCL_4693 Antigen Expression: cytokeratin 8, cytokeratin 19 Tumorigenic: No, No not tumorgenic in nude mice Viruses: SV40 Karyotype: Cells are reported to remain near diploid and karyotypically stable from passage 19-70 with 50% or more of the cells containing 42 chromosomes. At passage 37, there was a translocation between chromosome 1 and 15 which results in trisomy of the q arm of chromosome 1. Culture Medium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) Supplements: Supplement the medium with 5% FBS Dissociation Reagent: Accutase Fluid Renewal: 2 to 3 times per week Freeze Medium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. Thawing And Culturing Cells: Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks, for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes. Sterility: Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 1: 820400a Required Product 3: 860015.0 Required Product 4: 830100.0