Wilms1 Cells

In vitro cell culture model. Biochemical studies

The Wilms1 cell line was derived from a primary Wilms tumor sample obtained from a patient presenting with large bilateral kidney tumors, indicative of Wilms tumor, a pediatric nephroblastoma. This cell line harbors a homozygous nonsense mutation in the WT1 gene (c.149 C>A, p.S50X), which results in a truncated and non-functional WT1 protein. The WT1 gene, critical for kidney development and function, is frequently mutated in Wilms tumor, particularly in those with a stromal subtype that exhibits ectopic mesenchymal differentiation. Wilms1 cells, therefore, represent a unique in vitro model for studying the consequences of WT1 loss of function in tumor biology.
The Wilms1 cell line maintains a stable karyotype with no significant chromosomal abnormalities, allowing for reliable long-term culture. These cells exhibit a mesenchymal phenotype, characterized by the expression of vimentin and the absence of epithelial markers such as cytokeratin, consistent with their stromal origin. Additionally, the cell line demonstrates limited but notable mesenchymal differentiation capacity, including the ability to differentiate into muscle-like cells under appropriate conditions. This makes Wilms1 an invaluable tool for investigating the molecular mechanisms of mesenchymal differentiation and its deregulation in Wilms tumor pathogenesis.
Wilms1 has also been used to study the activation status of key signaling pathways involved in tumor progression. Proteomic analyses have shown that Wilms1 cells exhibit phosphorylation and activation of several receptor tyrosine kinases, including EGFR and PDGFRβ, as well as downstream MAPK signaling pathways. These findings highlight the relevance of the Wilms1 cell line in exploring targeted therapeutic approaches for Wilms tumor by dissecting the role of these pathways in cancer cell survival, proliferation, and differentiation.

Adherent

2 years

Caucasian

1

MSCGM kit (from Lonza)

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.

Fast

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.

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.

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.

Yes, in nude mice. Forms tumor with small cells consistent with Wilms tumor (xenografts may not represent Wilm?s tumors completely, see E. Kunce Stroup 2017)

24 hours

46, normal

Accutase