imWilms1 Cells




















General information
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, PDGFRβ, 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. |
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Organism | Human |
Tissue | Kidney |
Disease | Wilms Tumor |
Synonyms | IM-WT-1 |
Characteristics
Age | 10 months |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Spindle-shaped |
Cell type | Wilms cells |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | imWilms1 (Cytion catalog number 300412) |
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Biosafety level | 1 |
Expression / Mutation
Mutational profile | WT1 mutation status: homozygous c. 149 C>A, p.S50x, LOH: 11p11-11pter, CTNNB1 mutation status: heterozygous TCT>TTT, p.S45F |
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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. |
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. |
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,x
CSF1PO: 10,12
D13S317: 11,13
D16S539: 11,14
D5S818: 12,13,14
D7S820: 9,14
TH01: 9.3
TPOX: 8,9
vWA: 14,19
D3S1358: 14,17,18
D21S11: 30,31
D18S51: 15,18
Penta E: 5,14
Penta D: 13
D8S1179: 12,14
FGA: 22,25
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HLA alleles |
A*: '03:01:01, '24:02:01
B*: '35:03:01, '38:01:01
C*: '12:03:01
DRB1*: '07:01:01, '14:54:01
DQA1*: '01:04:01, '02:01:01
DQB1*: '02:02:01, '05:03:01
DPB1*: '02:01:02G, '04:02:01G
E: '01:03:01, '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.
Certificate of Analysis (CoA)
Lot Number | Certificate Type | Date | Catalog Number |
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300412-221 | Certificate of Analysis | 15. Apr. 2025 | 300412 |