Cat.No. : CSC-RO0275 Host Cell: Ba/F3
Size: >1x10^6 frozen cells/vial, 1 mL Stability: Stable in culture over a minimum of 10 passages
| Cat. No. | CSC-RO0275 |
| Description | This cell line is engineered to stably overexpress exogenous human FGFR4 bearing N535K mutation. |
| Target Gene | FGFR4 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | Ba/F3 |
| Host Cell Species | Mus musculus (Mouse) |
| Stability | Stable in culture over a minimum of 10 passages |
| Application | Drug screening and biological assays |
| Growth Conditions | 37 °C, 5% CO2 |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Size | >1x10^6 frozen cells/vial, 1 mL |
| Biosafety Level | 2 |
| Thawing & Subculturing Instructions | 1. Thaw cells by gently swirling in a 37°C water bath. To limit contamination, do not submerge the O-ring and cap. 2. When cells are ~70% thawed (~1 min), transfer the vial into a biosafety cabinet, and wipe the surface with 70% ethanol. Allow tube to dry completely. 3. Transfer the cells gently into a 15 mL conical tube containing 10 mL of pre-warmed culture medium (without antibiotic selection marker). Centrifuge cells at ~125 x g for 5~7 min. 4. Remove supernatant without disturbing the pellet, and resuspend cells in 1 mL culture medium (without antibiotic selection marker). Transfer cells to a 6-well plate containing ~2 mL pre-warmed growth medium (without antibiotic selection marker) or a T25 flask containing 5 mL pre-warmed culture medium (without antibiotic selection marker). 5. Incubate the culture at 37°C with 5% CO2. 6. Subculture: split saturated culture 1:4 ~ 1:6 every 3 days; seed out at about 1~3 x 10^5 cells/mL. |
| Growth Properties | Suspension, round |
| Freeze Medium | Frozen with 70% medium, 20% FBS, 10% DMSO |
| Freezing Instructions | Cells are recommended to generate additional frozen stocks at early passages. Frozen stocks should be preserved in a designated cryopreservation medium or in 70% RPMI 1640 + 20% FBS + 10% DMSO (without antibiotic selection marker). 1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO, without antibiotic selection marker) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250 x g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3 x10^6 cells/ml in chilled freezing medium. 6. Aliquot 1 ml of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
Rhabdomyosarcoma (RMS) is the most prevalent soft tissue sarcoma in children, characterized by its aggressive nature and poor prognosis, particularly in cases of relapse or metastasis. Central to its pathophysiology is the Fibroblast Growth Factor Receptor 4 (FGFR4), a receptor tyrosine kinase that plays a crucial role in muscle development and regeneration but is typically not expressed in mature muscle tissue. The aberrant amplification and activation of FGFR4 in RMS tumors have been linked to tumor progression, making FGFR4 a promising target for therapeutic intervention. In this context, researchers employed a chimeric Ba/F3 TEL-FGFR4 construct to evaluate the efficacy of several tyrosine kinase inhibitors, discovering that ponatinib (AP24534) exhibited significant inhibitory effects on RMS cells harboring either wild-type or mutated FGFR4.
Figure 1. The researchers utilized ponatinib to treat tumors with FGFR4 N535K and V550E mutations, demonstrating significant tumor growth inhibition, while tumors with wild-type FGFR4 showed no effect. (Li SQ, et al., 2013)
Creative Biogene's FGFR4-N535K Stable Cell Line - BaF3 provides a valuable resource for studying the role of FGFR4 in RMS and testing potential therapeutic agents like ponatinib. This stable cell line model is engineered to express the N535K mutation, which enhances sensitivity to FGFR4 inhibitors.
A: The Human FGFR4-N535K Stable Cell Line - BaF3 is uniquely engineered to express the N535K mutation in FGFR4, a variant significantly associated with several types of cancer. This specific mutation allows researchers to directly investigate the altered signaling pathways, cellular behaviors, and drug responses that are characteristic of cancer cells harboring this mutation, offering a precise model to study the role of FGFR4 in tumorigenesis and the development of targeted therapies.
A: Researchers can employ this cell line in high-throughput drug screening assays to identify compounds that inhibit FGFR4 signaling, particularly those that target the N535K mutation. By comparing the viability, proliferation, and signaling pathway activation of cells treated with various compounds to untreated controls, scientists can pinpoint substances that specifically antagonize the mutated FGFR4 function, thereby revealing potential therapeutic agents for cancers driven by this genetic alteration.
A: Culturing this cell line requires strict adherence to sterile techniques to prevent contamination. The culture medium should be supplemented with the appropriate selective agents to ensure the maintenance of the FGFR4-N535K mutation. Regular mycoplasma testing is recommended to avoid false results. Additionally, validating the expression of the mutant FGFR4 protein periodically through Western blotting or flow cytometry ensures that the cell line retains its critical phenotype for reliable experimental outcomes.
A: This cell line is invaluable in oncological research focused on cancers where FGFR4 mutations, especially the N535K variant, play a pivotal role in disease progression, such as rhabdomyosarcoma, breast, and liver cancers. It facilitates the study of mutant FGFR4-driven tumorigenesis, the examination of the molecular mechanisms underlying these cancers, and the assessment of the therapeutic potential of FGFR4 inhibitors, contributing to the development of targeted cancer treatments.
A: Effective methodologies include cell viability assays, such as MTT or CellTiter-Glo, to evaluate the cytotoxic effects of therapeutics. Phosphorylation assays can be used to assess the inhibition of FGFR4 signaling pathways. Additionally, apoptosis assays and cell cycle analysis can determine the mechanism of action of novel compounds. For a comprehensive evaluation, combining these assays with gene expression analysis through qRT-PCR or RNA sequencing can elucidate the broader impacts of therapeutic intervention on the cell line.
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
The Human FGFR4-N535K Stable Cell Line - BaF3 harbors the N535K mutation in FGFR4, allowing precise genetic modeling of this specific mutation's effects on cell behavior. This precision is critical for understanding mutation-specific cellular mechanisms and potential therapeutic intervention points.
This cell line ensures consistent expression of the mutated FGFR4, providing reliable and stable protein levels for experimentation. The Human FGFR4-N535K Stable Cell Line - BaF3's consistent protein expression is essential for conducting controlled and reproducible biological assays.
The BaF3 cell line is known for its robust growth and high viability, and the Human FGFR4-N535K Stable Cell Line - BaF3 maintains these characteristics. This high viability facilitates extended experimental procedures without the loss of cell integrity, allowing for long-term studies.
Due to the stable nature of the mutation and cell line, the Human FGFR4-N535K Stable Cell Line - BaF3 enables efficient data collection, reducing variability and enhancing the reliability of experimental results. This efficiency is crucial for generating high-quality data in research settings.
Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.
Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Copyright © Creative Biogene. All rights reserved.
