Cat.No. : CSC-RO0362 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-RO0362 |
| Description | This cell line is engineered to stably overexpress exogenous human ETV6-NTRK2 fusion protein bearing G639R mutation in NTRK2 part. |
| Target Gene | ETV6-NTRK2 |
| 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 |
Neurotrophic tropomyosin receptor kinase (NTRK) gene fusions are well-established oncogenic drivers in various cancers, including rare pediatric and adult malignancies. TRK inhibitors, developed to target these fusions, have significantly improved outcomes in NTRK-fusion cancers. However, resistance to first-generation inhibitors often emerges due to mutations such as G639R in NTRK2, which alters drug binding. Researchers have explored second-generation TRK inhibitors like BPI-28592, demonstrating efficacy in overcoming resistant mutations through its targeted binding and inhibition of TRK signaling. This work highlights the importance of stable cell lines, such as Ba/F3 cells expressing mutated forms of NTRK, in investigating the mechanisms of resistance and testing potential therapeutic candidates. These cell lines provide a robust platform to simulate in vivo conditions and assess drug responses in cancer research.
Figure 1. The table shows the proliferative inhibition effects of BPI-28592 on various cell lines with TRK fusions. (Sheng J, et al., 2024)
Creative Biogene offers the Human ETV6-NTRK2-G639R Stable Cell Line - BaF3, a reliable tool for similar studies on TRK inhibitor resistance. This cell line supports research on TRK-fusion cancers by enabling detailed investigation into the function of resistant mutations, providing valuable insights for drug development and therapeutic strategies.
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After multiple passages, key indicators such as cell growth characteristics, gene expression, and signal transduction have remained stable, greatly reducing experimental errors.
The cell lines cultivated from this product are compatible with various detection and phenotype analysis techniques, enhancing their practicality and research potential.
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