Cat.No. : CSC-RO0136 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-RO0136 |
| Description | Ba/F3-EGFR-L858R cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with L858R 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 |
Lung adenocarcinoma frequently harbors mutations in the epidermal growth factor receptor (EGFR) kinase domain, which are critical driver of oncogenesis. Researchers have investigated the unique characteristics of these mutations, particularly focusing on the L858R mutant, which requires asymmetric dimerization for its activation. This dimerization is essential for oncogenic transformation, unlike other mutants such as Ex19Del, Ex20Ins, and L858R/T790M, which demonstrate dimerization-independent activation. Notably, the effectiveness of cetuximab, an EGFR-targeting monoclonal antibody, varies depending on the dimerization dependency of the EGFR mutants, highlighting the need for tailored therapeutic approaches in treating resistant lung cancers.
Figure 1. The researchers employed Ba/F3 cells transformed with various EGFR mutants, treating them with cetuximab to assess growth suppression and receptor degradation effects. (Cho J, et al., 2013)
Creative Biogene's Human EGFR-L858R Stable Cell Line-Ba/F3 is a valuable tool for exploring these therapeutic avenues. This cell line can serve as a model for investigating the mechanisms of resistance in lung cancer therapies and the efficacy of novel treatments targeting EGFR mutations.
A: The Human EGFR-L858R Stable Cell Line-Ba/F3 provides a controlled cellular environment to study the oncogenic effects of the EGFR L858R mutation. By observing the behavior of these cells in vitro, researchers can gain insights into how this specific mutation leads to uncontrolled cell growth and proliferation, a hallmark of cancer. This model allows for the assessment of signaling pathways activated by the mutated EGFR and the downstream effects on cell survival, migration, and differentiation, all of which are critical in understanding oncogenesis.
A: The Human EGFR-L858R Stable Cell Line-Ba/F3 is an excellent model for screening and evaluating the efficacy of new EGFR TKIs. By treating these cells with various concentrations of TKIs, researchers can assess the compounds' ability to inhibit EGFR activity, thereby observing the impact on cell viability, proliferation, and apoptosis. This preclinical testing is crucial for the development of targeted therapies that can effectively treat EGFR-mutated cancers, such as NSCLC.
A: The Human EGFR-L858R Stable Cell Line-Ba/F3 is distinguished by its stable integration of the L858R mutation in the EGFR gene, which results in a specific activation of downstream signaling pathways. This cell line exhibits increased dependency on EGFR signaling for survival and growth, making it hypersensitive to EGFR inhibition. Additionally, the Ba/F3 background of this cell line provides a unique context for studying the L858R mutation, as it is a hematopoietic cell line that can be easily genetically manipulated, unlike some other cancer cell lines.
A: The Human EGFR-L858R Stable Cell Line-Ba/F3 is instrumental in studying resistance to first-generation EGFR TKIs. By culturing these cells with continuous exposure to these inhibitors, researchers can identify secondary mutations and bypass mechanisms that lead to resistance. Understanding these mechanisms is crucial for the design of second and third-generation EGFR inhibitors that can overcome resistance and provide more effective treatment options for patients with EGFR-mutated cancers.
A: Absolutely, the Human EGFR-L858R Stable Cell Line-Ba/F3 is well-suited for investigating the interplay between EGFR signaling and other cellular pathways, such as PI3K/AKT/mTOR and RAS/RAF/MEK. By modulating the activity of these pathways in conjunction with EGFR inhibition, researchers can assess the potential synergistic effects on cell growth and survival. This knowledge is vital for the rational design of combination therapies aimed at enhancing the efficacy of cancer treatments and minimizing the risk of resistance.
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This cell line is optimized for testing oncological responses, specifically how cells with the EGFR-L858R mutation respond to various therapies. The Human EGFR-L858R Stable Cell Line - Ba/F3 provides a direct approach to assess the efficacy of treatments aimed at cancers with this mutation.
We can utilize the Human EGFR-L858R Stable Cell Line - Ba/F3 to conduct drug sensitivity analysis, identifying how cancer cells harboring the L858R mutation react to different chemotherapeutic agents. This analysis is crucial for developing personalized medicine strategies for lung cancer patients.
The cell line aids in the development of personalized therapies by allowing us to test drugs on a specific, clinically relevant mutation. The Human EGFR-L858R Stable Cell Line - Ba/F3 helps in tailoring treatments that are most likely to be effective for patients with the L858R EGFR mutation.
The stable expression of the L858R mutation in the Human EGFR-L858R Stable Cell Line - Ba/F3 allows for detailed analysis of the molecular pathways affected by this mutation. We can investigate how the mutation alters EGFR signaling and promotes oncogenic processes, providing insights critical for effective drug design.
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