Wilms2 Cells




















General information
Description | 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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Organism | Human |
Tissue | Kidney |
Disease | Wilms tumor |
Applications | In vitro cell culture model. Biochemical studies |
Characteristics
Age | 1 year |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Spindle-shaped |
Cell type | Wilms cells |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | Wilms2 (Cytion catalog number 300413) |
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Biosafety level | 1 |
Expression / Mutation
Mutational profile | WT1 mutation status: homozygous c.149 C>A, p.R326x, LOH: 11p11-11pter, CTNNB1 mutation status: heterozygous del TCT>TAT, p.S45Y |
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Handling
Culture Medium | MSCGM kit (from Lonza) |
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Passaging solution | 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. |
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. |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
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. |
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STR profile |
Amelogenin: x,y
CSF1PO: 11,11
D13S317: 9,11
D16S539: 9,9
D5S818: 11,11
D7S820: 10,11
TH01: 6,6
TPOX: 8,11
vWA: 15,19
D3S1358: 15,15
D21S11: 29,32.2
D18S51: 12,17
Penta E: 11,15
Penta D: 10,12
D8S1179: 14,16
FGA: 21,21
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HLA alleles |
A*: '01:01:01, '02:01:01
B*: '15:01:01, '57:01:01
C*: '03:03:01, '07:01:01
DRB1*: '04:01:01, '07:01:01
DQA1*: '02:01:01, '03:01:01
DQB1*: '03:02:01, '03:03:02
DPB1*: '04:01:01G, '04:02:01G
E: '01:01:01, '01:03:02
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Required products
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a widely used buffer solution in biological and chemical research. It plays a crucial role in maintaining the pH balance and osmolarity during various experimental procedures, including tissue processing and cell culture. Our PBS solution is meticulously formulated with high-purity ingredients to ensure stability and reliability in every experiment. The osmolarity and ion concentrations of our PBS closely mimic those of the human body, making it isotonic and non-toxic to most cells.
Composition of Our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains:
8000 mg/L Sodium chloride (NaCl)
200 mg/L Potassium chloride (KCl)
1150 mg/L Sodium phosphate dibasic anhydrous (Na2HPO4)
200 mg/L Potassium phosphate monobasic anhydrous (KH2PO4)
This composition ensures an optimal pH and ionic balance, suitable for a wide range of biological applications.
Applications of Our PBS Solution
Our PBS solution is ideal for various applications in biological research. Its isotonic and non-toxic properties make it suitable for substance dilution and cell container rinsing. PBS solutions containing EDTA are effective for disengaging attached and clumped cells. However, divalent metals such as zinc should not be added to PBS, as this can cause precipitation. In such cases, Good's buffers are recommended. Additionally, our PBS solution is an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, including SARS-CoV-2.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +25°C, protected from light.
Once opened, store at 2°C to 25°C and use within 24 months.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Salts
Potassium chloride
200
Potassium phosphate monobasic anhydrous
200
Sodium chloride
8000
Sodium phosphate dibasic anhydrous
1150