HCE-T Cells

Categories: Transformed cell lines Description: The HCE-T cell line is derived from human corneal epithelial cells and has been immortalized by transfection with the plasmid RSV-T (pRSV-T). This plasmid carries an SV40 origin of replication (SV40 ori) and includes the SV40 early region genes along with the Rous sarcoma virus long terminal repeat (RSV-LTR). This genetic modification enables the HCE-T cells to bypass normal senescence, allowing for indefinite proliferation. Despite their immortalization, these cells retain many of the functional and phenotypic characteristics of primary corneal epithelial cells, including typical epithelial morphology, the ability to form adherent monolayers, and the expression of cytokeratins and other essential epithelial markers. As a result, HCE-T cells are an invaluable resource for in vitro ocular research. HCE-T cells are widely utilized to investigate various aspects of corneal epithelial biology, such as cell adhesion, migration, differentiation, and response to external stimuli, including therapeutic agents and environmental stressors. Their ability to mimic in vivo corneal conditions makes them highly relevant for studying corneal diseases like dry eye syndrome, keratitis, and corneal ulcers. These cells serve as an important model to explore disease mechanisms and test potential treatments. Additionally, HCE-T cells play a crucial role in drug development and testing for ophthalmic applications, offering a reliable platform to evaluate the safety and efficacy of new drugs and drug delivery systems. Their capacity for long-term culture allows for sustained research efforts, advancing the development of new therapies aimed at improving ocular health. Organism: Human Tissue: Eye, cornea, epithelium Synonyms: HCET, Human Corneal Epithelial cells-Transformed, HCE, SV40-HCEC Age: 49 years Gender: Female Ethnicity: Japanese Morphology: Epithelial Cell Type: Epithelial cell Growth Properties: Adherent Citation: HCE-T (Cytion catalog number 305255) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_1272 Viruses: Transformant: plasmid RSV-T (pRSV-T). This plasmid is an SV40 ori-construct containing the SV40 early region genes and the Rous sarcoma virus long terminal repeat. Products: Keratin (64kD) 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, 1% ITS (0.625 mg/mL human insulin, 0.625 mg/mL human transferrin, 0.625 microgram/mL sodium selenite, 0.535 mg/mL linoleic acid, 125 mg/mL BSA) and 10 ng/mL human EGF 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. 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