imWilms1 Cells

Categories: Kidney cancer cell lines Description: The Wilms1 cell line was originally derived from a primary Wilms tumor, obtained from a patient diagnosed with large bilateral kidney tumors, a characteristic presentation of Wilms tumor (nephroblastoma). This cell line harbors a homozygous nonsense mutation in the WT1 gene (c.149 C>A, p.S50X), leading to the production of a truncated, non-functional WT1 protein. WT1 is a critical gene in kidney development, and its mutation is closely associated with the pathogenesis of Wilms tumor, particularly in tumors exhibiting stromal differentiation. Wilms1 cells display a stable karyotype without significant chromosomal abnormalities, and they are characterized by a mesenchymal phenotype, expressing vimentin while lacking epithelial markers like cytokeratin. The line shows a limited but significant capacity for mesenchymal differentiation, including the potential to differentiate into muscle-like cells under specific conditions, making it a crucial model for studying the molecular consequences of WT1 mutations. To overcome the limited lifespan of the primary Wilms1 cells, the imWilms1 cell line was established by introducing a triple mutant SV40 large T antigen (U19dl89-97tsA58) into the original tumor cells, facilitating their immortalization. This modification allows imWilms1 cells to proliferate indefinitely while maintaining chromosomal stability, thereby offering a reliable model for long-term studies. The immortalized imWilms1 cells continue to exhibit the same WT1 mutation and retain the mesenchymal characteristics of the parent Wilms1 line. In addition to its genetic and phenotypic features, the imWilms1 cell line has been extensively analyzed for its signaling pathway activity. Proteomic studies have revealed the phosphorylation and activation of several receptor tyrosine kinases (RTKs), including EGFR, PDGFRbeta, and AXL, with downstream activation of the MAPK signaling pathways. The consistent activation of these pathways in imWilms1 cells underscores their relevance for exploring targeted therapeutic strategies in Wilms tumor. Overall, imWilms1 serves as a robust and long-term model for investigating the molecular mechanisms underlying Wilms tumor development and progression, particularly those driven by WT1 mutations and aberrant signaling pathways. Organism: Human Tissue: Kidney Disease: Wilms Tumor Synonyms: IM-WT-1 Age: 10 months Gender: Female Ethnicity: Caucasian Morphology: Spindle-shaped Cell Type: Wilms cells Growth Properties: Adherent Citation: imWilms1 (Cytion catalog number 300412) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Cellosaurus Accession: CVCL_A5SN Mutational Profile: WT1 mutation status: homozygous c. 149 C>A, p.S50x, LOH: 11p11-11pter, CTNNB1 mutation status: heterozygous TCT>TTT, p.S45F Culture Medium: MSCGM kit (from Lonza) 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: 1 to 2 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 3: 860015.0 Required Product 4: 830100.0