Cat.No. : CSC-RO0137 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-RO0137 |
| Description | Ba/F3-EGFR-L858R/C797S cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with L858R and C797S mutations. |
| 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 |
Research into resistance mechanisms for third-generation EGFR inhibitors, such as osimertinib, highlights the significant challenge posed by mutations like C797S in EGFR. This mutation limits treatment options for patients, as no targeted therapies are currently available to address it effectively. In a recent study, BBT-176, a novel EGFR tyrosine kinase inhibitor, demonstrated potent inhibition against various mutant forms of EGFR, including the L858R/C797S variant. The findings indicated that BBT-176 achieved lower IC50 values than osimertinib, showcasing its potential as a more effective option for overcoming resistance associated with these mutations. The study also suggested that combining BBT-176 with the anti-EGFR antibody cetuximab could enhance its efficacy, particularly against resistant mutant cells.
Figure 1. The researchers employed ENU mutagenesis to derive BBT-176-resistant clones from Ba/F3 cells, assessing their growth inhibition and IC50 values to understand resistance mechanisms. (Lim SM, et al., 2023)
Creative Biogene's Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 cells provide a powerful tool for researchers investigating the therapeutic effects of novel inhibitors targeting mutant EGFR variants. This cell line can be utilized in high-throughput screening to assess the efficacy of new compounds, including BBT-176 and other emerging therapies.
A: The Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 achieves stable expression of specific EGFR mutations by introducing the mutated EGFR gene carrying the L858R and C797S mutations into the Ba/F3 cell line using genetic engineering techniques. This typically involves the use of retroviral or lentiviral vectors to integrate the mutated gene into the host cell genome, and antibiotic selection to ensure that only cells containing the mutated gene survive and proliferate, thus achieving stable expression of the mutated EGFR.
A: The role of the Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 in studying EGFR inhibitor resistance is to provide a model system for simulating and investigating how EGFR mutations lead to resistance to existing therapeutic drugs. This cell line helps to uncover resistance mechanisms and provides a scientific basis for developing new treatment strategies.
A: The Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 assists in drug screening by providing a cellular model with specific EGFR mutations to test and screen for new EGFR inhibitors. This cell line helps scientists evaluate the inhibitory effects of new compounds on mutated EGFR, thereby screening for potentially effective drug candidates.
A: The Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 models EGFR mutations in lung cancer by stably expressing the EGFR gene with L858R and C797S mutations in the Ba/F3 cell line. This cell line allows researchers to observe how these mutations affect cellular behavior, especially in terms of resistance and signaling, which is crucial for developing targeted therapies for these mutations.
A: The role of the Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 in studying the tumor microenvironment in lung cancer is to simulate the behavior of lung cancer cells within the microenvironment, investigating how EGFR mutations affect cell interactions with the surrounding environment. This cell line helps understand cellular signaling and interactions within the tumor microenvironment, which is significant for developing treatment strategies targeting the tumor microenvironment.
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The Human EGFR-L858R/C797S Stable Cell Line-Ba/F3 offers stable expression suitable for long-term experiments and studies.
This product ensures reliable data, guaranteeing accuracy and reproducibility of experimental results.
The high expression level of EGFR-L858R/C797S in Ba/F3 cell line facilitates the detection and study of its biological functions.
With this stable cell line, you can initiate experiments rapidly, saving time and resources.
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