Caki-2 Cells

Categories: Kidney cancer cell lines Description: Caki-2 is a human clear cell renal cell carcinoma (ccRCC) cell line that exhibits epithelial morphology and adheres during in vitro culture conditions. It serves as an essential preclinical model for the investigation of renal cancer mechanisms and therapeutic responses. The Caki-2 line is particularly notable for its resistance to certain chemotherapeutic agents, it displays decreased sensitivity to 5-fluorouracil and the multi-kinase inhibitor sorafenib, which targets VEGFRs 1-3, PDGFR-b, and Raf-1, in comparison to the Caki-1 cell line. This differential sensitivity is significant for studying drug resistance mechanisms and evaluating new therapeutic strategies in renal cell carcinoma. The genetic background of Caki-2 cells includes a loss-of-function mutation in the von Hippel-Lindau (VHL) tumor suppressor protein, a hallmark of many ccRCCs that leads to the deregulation of hypoxia-inducible factors (HIFs) and contributes to tumorigenesis. The ability of Caki-2 cells to form tumors in immunocompromised mice makes them a valuable tool for in vivo studies of cancer growth and metastasis, providing insights into the tumor environment and potential therapeutic interventions. Their use extends to exploring the role of VHL in cancer progression and testing the efficacy of drugs targeting the HIF pathway and other associated signaling cascades in a controlled experimental setup. Organism: Human Tissue: Kidney Disease: Papillary carcinoma Synonyms: CAKI-2, CaKi-2, caki-2, CAKI 2, Caki 2, Caki2, CAKI2 Age: 69 years Gender: Male Ethnicity: Caucasian Morphology: Epithelial-like. Ultrastructural features include microvilli and microfilaments. Few mitochondria, lysosomes or lipid droplets. Frequent multilamellar bodies. No virus particles. Growth Properties: Monolayer, adherent Citation: Caki-2 (Cytion catalog number 300140) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_0235 Isoenzymes: Me-2, 1, PGM3, 1, PGM1, 1, ES-D, 1, AK-1, 1, GLO-1, 1-2, G6PD, B, Phenotype Frequency Product: 0.0511 Tumorigenic: Yes, in nude mice. Forms clear cell carcinoma Karyotype: (P8) hypopentaploid to hypohexaploid (+A2, +A3, +B, +C, +D, +F, +G, -A) with abnormalities including dicentrics, acrocentric fragments, minutes, breaks, and large subtelocentric markers Culture Medium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) Supplements: Supplement the medium with 10% FBS 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. Seeding Density: 1 x 104 cells/cm2 will result in a 90% confluent monolayer in about 4 days Fluid Renewal: 2 to 3 times per week Post Thaw Recovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. 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: 820700a Required Product 3: 860015.0 Required Product 4: 830100.0