Cat.No. : CSC-RO0127 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-RO0127 |
| Description | Ba/F3-EGFR-C797S cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with C797S mutation. |
| Target Gene | EGFR |
| 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 |
| 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 |
EGFR (Epidermal Growth Factor Receptor) is a critical oncogene implicated in the progression of non-small cell lung cancer (NSCLC). Mutations in the EGFR gene, such as exon 20 insertions or resistance mutations like EGFR-C797S, are associated with treatment resistance to tyrosine kinase inhibitors (TKIs). Researchers use stable cell lines to investigate the mechanisms of drug resistance and evaluate the efficacy of next-generation inhibitors targeting these mutations. In this context, Ba/F3 murine pro-B lymphocyte cells, engineered to express EGFR mutants, have become essential for preclinical studies. They allow scientists to model EGFR-driven oncogenesis and test the therapeutic windows of various inhibitors.
Figure 1. The researchers utilized Ba/F3 cell lines harboring EGFR mutations, such as T790M and C797S, to investigate how these mutations affect the sensitivity of various EGFR TKIs. (Kobayashi IS, et al., 2023)
Creative Biogene's Human EGFR-C797S Stable Cell Line-Ba/F3 offers a robust platform for similar research, particularly for studying resistance mutations like C797S. This stable cell line can be used to evaluate the efficacy of covalent and non-covalent TKIs, facilitating the development of novel therapeutic strategies against drug-resistant lung cancers.
A: The Human EGFR-C797S Stable Cell Line-Ba/F3 is particularly suitable for applications in drug discovery and development, especially for the identification and evaluation of new therapeutic agents targeting the EGFR-C797S mutation. This cell line can also be used in studies focused on understanding the molecular mechanisms underlying resistance to EGFR tyrosine kinase inhibitors (TKIs) in lung cancer, as well as in functional assays to screen for compounds that can overcome this resistance.
A: The EGFR-C797S mutation confers resistance to third-generation EGFR tyrosine kinase inhibitors (TKIs) such as osimertinib, which are designed to target T790M mutation-positive cancers. The use of the Human EGFR-C797S Stable Cell Line-Ba/F3 allows researchers to study the effectiveness of novel TKIs or combination therapies in overcoming this resistance, as the mutation affects the binding site of these inhibitors, rendering them less effective.
A: Yes, the Human EGFR-C797S Stable Cell Line-Ba/F3 is an excellent tool for investigating the downstream signaling pathways affected by the EGFR-C797S mutation. Researchers can use this cell line to analyze changes in signaling pathways such as PI3K/AKT and MAPK, which are crucial for cell proliferation, survival, and apoptosis. This can help in understanding the biological effects of the mutation and in identifying potential therapeutic targets to counteract the oncogenic signals.
A: When culturing the Human EGFR-C797S Stable Cell Line-Ba/F3, it is essential to maintain optimal growth conditions such as appropriate temperature, CO2 levels, and nutrient-rich media specific to Ba/F3 cells. Regular monitoring for mycoplasma contamination and genetic drift is crucial. Additionally, selecting the correct antibiotic for maintaining selective pressure is necessary to ensure the stable expression of the EGFR-C797S mutation. Periodic genetic verification through sequencing or PCR may also be beneficial to confirm the integrity of the mutation.
A: One potential challenge in using the Human EGFR-C797S Stable Cell Line-Ba/F3 in high-throughput screening (HTS) assays is the need to ensure uniform cell growth and consistent expression of the EGFR-C797S mutation across all wells, which is crucial for reliable results. Additionally, the specific sensitivity of the cell line to various compounds may necessitate the optimization of assay conditions, including drug concentration and incubation times. Overcoming these challenges requires careful planning and pilot studies to establish optimal assay conditions.
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This cell line allows for detailed genetic studies focusing on the impact of the EGFR-C797S mutation on cell signaling and behavior. The Human EGFR-C797S Stable Cell Line - Ba/F3 helps us understand how this mutation alters EGFR function and contributes to disease progression.
We can use the Human EGFR-C797S Stable Cell Line - Ba/F3 to investigate the mechanisms by which cancer cells develop resistance to current EGFR inhibitors. Understanding these mechanisms is essential for the development of next-generation therapeutics designed to be effective against resistant strains.
This cell line is particularly valuable for oncology research, facilitating the study of tumor cell behavior and drug resistance in cancers that exhibit the EGFR-C797S mutation, such as non-small cell lung cancer. The Human EGFR-C797S Stable Cell Line - Ba/F3 provides a relevant model to explore therapeutic strategies
The Human EGFR-C797S Stable Cell Line - Ba/F3 contains a specific mutation (C797S) in the EGFR gene, crucial for studying resistance mechanisms against EGFR inhibitors used in cancer treatment. This specific focus allows us to better understand and address resistance in therapies targeting EGFR mutations.
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