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ImWilms10T Cells Europäischer Partner

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ArtNr CLS-300419
Hersteller CLS Cell Lines Service
Menge 1 cryovial
Kategorie
Typ Cell line
Zertifikat For a certificate of analysis, please send an inquiry to info@hoelzel.de.
Specific against Human (Homo sapiens)
Dry ice Yes
ECLASS 10.1 42040401
ECLASS 11.0 42040401
UNSPSC 41106509
Alias ImWilms10 T,IM-WT-10
Lieferbar
Manufacturer - Category
Kidney cancer cell lines
Description
The imWilms10T cell line is an immortalized variant of the Wilms10T primary tumor cell line, which was derived from a Wilms tumor (nephroblastoma) sample in a pediatric patient. This cell line is distinguished by a homozygous deletion of the WT1 gene, resulting in a complete loss of WT1 protein function. WT1 is a crucial gene for kidney development, and its deletion in imWilms10T reflects a severe genetic disruption that is associated with the pathogenesis of Wilms tumor. In addition to the WT1 deletion, imWilms10T cells exhibit loss of heterozygosity (LOH) in the 11p15 chromosomal region, which includes key genes such as IGF2, contributing to the tumor's aggressive behavior.

To overcome the limited lifespan of the Wilms10T cells, the imWilms10T cell line was established by introducing a triple mutant SV40 large T antigen (U19dl89-97tsA58) into the original tumor cells. This immortalization process enables imWilms10T cells to proliferate indefinitely while maintaining chromosomal stability, thereby providing a reliable model for long-term studies. The imWilms10T cells retain the critical characteristics of the parental Wilms10T line, including the complete loss of WT1 and the presence of LOH at 11p15, making them an invaluable resource for studying the molecular consequences of WT1 deletion and the associated tumorigenic processes.

imWilms10T cells have been extensively studied for their involvement in key signaling pathways that drive tumor progression. Proteomic analyses have revealed that these cells exhibit phosphorylation and activation of several receptor tyrosine kinases (RTKs), such as IGF1R, PDGFRβ, and AXL. These activated receptors signal through downstream pathways, including the MAPK and PI3K/AKT pathways, which are crucial for maintaining the malignant phenotype of the cells. The imWilms10T cell line serves as an important tool for investigating the impact of complete WT1 loss on cellular signaling, tumor growth, and potential therapeutic targets in Wilms tumor, particularly for more aggressive tumor subtypes.
Tissue
Kidney
Growth properties
Adherent
Cell type
Wilms cells
Disease
Wilms Tumor
Age
2 years
Gender
Female
Ethnicity
Caucasian
Morphology
Spindle-shaped
Biosafety Level
1
Culture Medium
MSCGM kit (from Lonza)
Medium Supplements
4 mM L-glutamine
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
Handling of Cryopreserved Cultures

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.

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.
Safety Precautions
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.
Disclaimer
Our cells are provided for in vitro laboratory research purposes exclusively and are not intended for clinical or diagnostic use, nor are they to be administered to humans or used for veterinary purposes. Users must adhere to all applicable guidelines and regulations for the handling and use of these cells in a research setting.
Warranty
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.
Amelogenin
X, X
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

Hinweis: Die dargestellten Informationen und Dokumente (Bedienungsanleitung, Produktdatenblatt, Sicherheitsdatenblatt und Analysezertifikat) entsprechen unserem letzten Update und sollten lediglich der Orientierung dienen. Wir übernehmen keine Garantie für die Aktualität. Für spezifische Anforderungen bitten wir Sie, uns eine Anfrage zu stellen.

Alle Produkte sind nur für Forschungszwecke bestimmt. Nicht für den menschlichen, tierärztlichen oder therapeutischen Gebrauch.

Menge: 1 cryovial
Lieferbar: In stock
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