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The Wilms10M cell line was established from a metastatic lung nodule of a patient with Wilms tumor (nephroblastoma). Like its primary tumor counterpart, Wilms10T, the Wilms10M cell line is characterized by a homozygous deletion of the WT1 gene, resulting in the complete absence of WT1 protein. WT1 is essential for normal kidney development, and its deletion is associated with more aggressive tumor behavior, particularly in metastatic settings. Additionally, Wilms10M cells exhibit loss of heterozygosity (LOH) in the 11p15 chromosomal region, which includes the IGF2 gene, further contributing to the malignant properties of these cells.
Wilms10M cells maintain a stable karyotype with no major chromosomal rearrangements apart from the specific deletion of the WT1 region. This cell line, derived from metastatic tissue, is particularly valuable for studying the molecular mechanisms that drive metastasis in Wilms tumor. The cells exhibit mesenchymal characteristics, expressing markers such as vimentin, while lacking epithelial markers like cytokeratin, which is indicative of their origin from the stromal component of the tumor.
Research on Wilms10M has focused on the signaling pathways that are active in these metastatic cells. Proteomic analyses have demonstrated the activation of several receptor tyrosine kinases (RTKs), including IGF1R, PDGFRβ, and AXL, which are involved in promoting cell survival, proliferation, and metastatic potential. The downstream MAPK and PI3K/AKT signaling pathways are also activated, playing a key role in maintaining the invasive and metastatic phenotype of Wilms10M cells. Given its metastatic origin, Wilms10M is an essential model for understanding the molecular events underlying Wilms tumor metastasis and for developing targeted therapeutic strategies against metastatic disease.
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Mutational profile: WT1 mutation status: homozygous del WT1 within del11p13. LOH: no in 11p13 but UPD in 11p15. CTNNB1 mutation status: homozygous del TCT, p.DS45, UPD 3p
- 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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