Cat.No. : CSC-RO0347 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-RO0347 |
| Description | This cell line is engineered to stably overexpress human C-Kit with V560G mutation. |
| Target Gene | KIT |
| 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 |
A: This cell line harbors the KIT-V560G mutation commonly found in GISTs, allowing for mechanistic studies of mutated KIT signaling pathways and cellular behaviors like proliferation and apoptosis in GISTs.
A: TKIs can be applied to the cell line, and their effects on cell viability, proliferation, and KIT signaling can be measured to assess the drugs' efficacy and potential resistance mechanisms.
A: Sequencing the KIT gene region of interest, along with protein expression analysis via Western blot or flow cytometry, is necessary to confirm the presence and stable expression of the KIT-V560G mutation.
A: The Human KIT-V560G Stable Cell Line - BaF3 is a valuable model for studying the pathophysiology of gastrointestinal stromal tumors (GISTs), as the KIT-V560G mutation is a common driver in GISTs. It allows researchers to examine the cellular signaling, growth characteristics, and response to therapy that this mutation confers to the cells.
A: The KIT-V560G mutation in the BaF3 cell line results in constitutive activation of the KIT receptor, which may lead to altered sensitivity to various tyrosine kinase inhibitors. This model enables the comparison of drug responses and the exploration of resistance mechanisms to treatments that are currently used in clinical settings for GISTs.
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Provides a relevant model for gastrointestinal stromal tumors (GIST), where KIT mutations are common, aiding in the understanding of disease mechanisms.
This cell line is engineered to carry the KIT-V560G mutation, enabling us to study the effects of this specific oncogenic mutation in detail.
Ideal for pharmacological studies due to the stable expression of the KIT-V560G protein, allowing for precise drug efficacy evaluations.
The Human KIT-V560G Stable Cell Line - BaF3 serves as a comprehensive tool for both basic research into the biology of KIT mutations and for translational research into potential treatments.
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