CTLA4 Ig-24 Cells

Categories: Hamster cell lines Description: CTLA4 Ig-24 cells, derived from an adult female Chinese hamster (Cricetulus griseus), are a spontaneously immortalized cell line which have been genetically modified by introducing the human CTLA-4 gene, resulting in the expression of a fusion protein. The fusion protein possesses a dominant attribute of CTLA4Ig, making CTLA4 Ig-24 cells a unique and essential tool in immunology research. CTLA-4, a member of the immunoglobulin superfamily, is primarily expressed in activated T cells and acts as an inhibitory signal transmitter to regulate T cell function. It shares homology with CD28, a T-cell co-stimulatory protein, and both molecules bind to CD80 (B7-1) and CD86 (B7-2) proteins on antigen-presenting cells. CTLA-4 demonstrates a greater affinity and avidity for CD80 and CD86 than CD28, allowing it to outcompete CD28 for binding to these ligands. By doing so, CTLA-4 transmits an inhibitory signal to T cells, while CD28 transmits a stimulatory signal. This intricate regulatory mechanism is pivotal in maintaining immune balance and preventing excessive immune responses. Interestingly, CTLA-4 is also found in regulatory T cells (Tregs) and contributes to their inhibitory function. When T cells are activated through the T cell receptor (TCR) and CD28, the expression of CTLA-4 increases. Furthermore, CTLA-4 may influence cell motility and signal signalling through the PI3 kinase pathway. Organism: Chinese hamster Tissue: Ovary Synonyms: CTLA4Ig-24 Gender: Female Morphology: Epithelial Growth Properties: Adherent Citation: CTLA4 Ig-24 (Cytion catalog number 305014) Biosafety Level: 1 Ncbi_ Taxid: 10029.0 Cellosaurus Accession: CVCL_6384 Culture Medium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) Supplements: Supplement the medium with 10% FBS, 0.2 mM proline, 0.001 mM methotrexate Dissociation Reagent: Accutase Subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. 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: 820300a Required Product 3: 860015.0 Required Product 4: 830100.0