SiHa Cells

Categories: Reproductive system cancer cell lines Description: SiHa cells are a human cervical squamous cell carcinoma cell line that has been widely used in research for several decades. They were isolated from primary uterine biopsy fragments from a 55-year-old female Japanese patient with squamous cell carcinoma. This cell line is of great interest to researchers studying cervical cancer and other related diseases due to their unique genetic characteristics. SiHa cells have been found to express the p53+ and pRB+ genes, which are involved in cell cycle regulation, DNA repair, and tumor suppression. These genes make SiHa cells an ideal model for studying the molecular mechanisms of cancer development and progression. Additionally, SiHa cells are a suitable transfection host, making them an excellent tool for gene expression studies. SiHa cells have a hypertriploid karyotype, with an average chromosome number between 69 and 72. The SiHa cells are HPV-16 positive, showing integration of 1 to 2 copies of the viral genome per cell. Cells are tumorigenic, forming poorly differentiated epidermoid carcinoma (grade III) in nude mice. This makes them an excellent model for studying cancer progression and testing anti-cancer drugs. SiHa cell line expresses various isoenzymes, including AK-1, ES-D, G6PD, GLO-I, Me-2, PGM1, and PGM3. Electron microscopy revealed abundant tonofilaments in the cytoplasm and desmosomes at the cell junctions. The growth properties of SiHa cells are adherent, with a doubling time of 17 hours in 10% FBS media and 21 hours in 5% FBS media. Epithelial cell adhesion molecule (EpCAM) expression is present in 92% of SiHa cells, indicating their epithelial origin. They show strong cytokeratin expression but no vimentin expression. Organism: Human Tissue: Cervix Disease: Human papillomavirus-related cervical squamous cell carcinoma Synonyms: Siha, SIHA Age: 55 years Gender: Female Ethnicity: Asian Morphology: Epithelial Growth Properties: Adherent Citation: SiHa (Cytion catalog number 305023) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_0032 Tumorigenic: Yes Culture Medium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a) Supplements: Supplement the medium with 10% FBS and 1% NEAA 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: 820100a Required Product 3: 860015.0 Required Product 4: 830100.0