| Field | Specification |
|---|---|
| Species |
The Wilms2 cell line was derived from a primary Wilms tumor in a pediatric patient with a germline WT1 mutation. This cell line is characterized by a homozygous nonsense mutation in the WT1 gene (c.1084 C>T, p.R362X), which results in the production of a truncated, non-functional WT1 protein. The loss of functional WT1, a gene essential for kidney development, is a hallmark of certain subtypes of Wilms tumor, particularly those associated with mesenchymal or stromal differentiation. The Wilms2 cell line is a significant model for studying the tumorigenic processes driven by WT1 loss, especially in the context of Wilms tumors that retain other critical genetic features.
Wilms2 cells also carry mutations in the CTNNB1 gene, which encodes β-Catenin, a key component of the Wnt signaling pathway. These mutations, specifically affecting serine 45, lead to the stabilization and accumulation of β-Catenin, resulting in the constitutive activation of the Wnt pathway. This activation is a known driver of cell proliferation and tumorigenesis in Wilms tumor, making Wilms2 a valuable model for understanding how aberrant Wnt signaling contributes to the development and progression of tumors with WT1 mutations.
In terms of phenotype, Wilms2 cells exhibit a mesenchymal-like morphology, expressing vimentin and lacking epithelial markers such as cytokeratin. This aligns with the tumor's stromal characteristics and underscores the role of WT1 in regulating mesenchymal-epithelial transitions during kidney development. Proteomic analyses of Wilms2 have identified activation of several receptor tyrosine kinases (RTKs), including PDGFRβ and AXL, which are known to support tumor cell survival and proliferation. Additionally, downstream pathways such as MAPK and PI3K/AKT are also activated, further contributing to the malignant properties of Wilms2 cells.
Overall, the Wilms2 cell line serves as an essential tool for exploring the molecular mechanisms of Wilms tumor driven by WT1 loss and aberrant Wnt signaling. Its genetic and phenotypic characteristics provide a robust platform for investigating potential therapeutic targets and for understanding the role of key signaling pathways in the pathology of Wilms tumors with a mesenchymal component.
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Mutational profile: WT1 mutation status: homozygous c.149 C>A, p.R326x, LOH: 11p11-11pter, CTNNB1 mutation status: heterozygous del TCT>TAT, p.S45Y
- cultureMedium: MSCGM kit (from Lonza)
- dissociationReagent: 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.
- freezeMedium: 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.
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