HEK293 suspension-adapted

Categories: Kidney cancer cell lines , Transformed cell lines Description: The HEK293 suspension-adapted cell line is a variant of the human embryonic kidney 293 (HEK293) cells that has been modified to grow in suspension culture rather than adherent culture. This adaptation is significant for industrial applications where large-scale protein production is required. The cells maintain many of the characteristics of the original HEK293 line, including a robust transient transfection efficiency and the ability to post-translationally modify expressed proteins in a manner similar to that of native human cells. These cells are particularly valued in the biotechnology and pharmaceutical industries for the production of recombinant proteins and viruses for gene therapy and vaccine development. The adaptation to suspension culture allows for easier scalability and simplifies the harvesting process, making it more suitable for commercial-scale bioprocessing. The HEK293 suspension-adapted cell line supports various viral production systems, including adenovirus, lentivirus, and adeno-associated virus (AAV), which are pivotal in therapeutic applications and research. Overall, the HEK293 suspension-adapted cell line is a crucial tool in the fields of molecular biology and bioprocessing, providing a versatile platform for the production of various biologically active molecules. Its ease of genetic manipulation and ability to produce proteins that are correctly folded and post-translationally modified according to human cell patterns make it an indispensable resource in many advanced therapeutic and research settings. Organism: Human Tissue: Kidney Age: Fetus Gender: Female Morphology: round Growth Properties: Suspension Citation: HEK293 suspension-adapted (Cytion catalog number 300686) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_0045 Receptors Expressed: Vitronectin Protein Expression: CEA negative, p53 positive Tumorigenic: In nude mice Virus Susceptibility: transformed with adenovirus 5 DNA adenovirus 5 DNA Culture Medium: Panserin 293S (PanBiotech, Germany) Supplements: No supplements required Dissociation Reagent: Not required Subculturing: Maintain the suspension cells at cell densities between 5 x 105 and 2-3 x 106 cells/ml in Eppendorf cell culture flasks on a shaker inside an incubator at 37°C/5% CO2. Subculture once the cell density has reached 2-3 x 106 cells/ml. Carefully dislodge the cells to avoid cluster. Seeding Density: 5 x 105 cells/ml Post Thaw Recovery: Initiate cultures at a density of 5 x 105 cells/ml and keep the cell concentration up to 2-3 x 106 cells/ml for optimal growth. Incubate at 37°C/5% CO2 on a cell shaker at 100-150 rpm. Freeze Medium: As a cryopreservation medium, use complete growth medium + 10% DMSO for adequate post-thaw viability. 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 200 x g for 5 minutes, carefully discard the supernatant containing freezing medium. Follow the procedure described under Post-Thaw Recovery 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. Freezing Procedure: Once the cell density of 1-2 x 106 cells/ml is achieved, collect the cells by ccentrifuging at 200xg for 5 min and discard the supernatant. Dilute in an appropriate volume of fresh, prewarmed culture medium and count the cells to get information on the viability and number of cells. Collect the cells by ccentrifuging at 200xg for 5 min and discard the supernatant. Resuspend the cells in the appropriate volume of freeze medium and count once more. The cell viability should be >>80%, a cell density of 5-10 million cells/ml is recommended. Pipette the cells into pre-labeled cryovials. Use either CoolCell freezing container or a controlled rate freezer to ensure a cooling rate of 1°C/min. 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 3: 860015.0