Cat.No. : CSC-RO0139 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-RO0139 |
| Description | Ba/F3-EGFR-L858R/T790M/C797S cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with L858R, T790M 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 |
First-, second-, and third-generation EGFR inhibitors have greatly prolonged survival in advanced EGFR-mutant NSCLC patients (refs. 1–10). Researchers have developed a Human EGFR-L858R/T790M/C797S Stable Cell Line to address resistance to third-generation EGFR inhibitors such as osimertinib, attributed in part to the C797S mutation in EGFR. BBT-176, a new EGFR tyrosine kinase inhibitor (TKI) targeting the C797S mutation, was developed for these patients lacking targeted treatment options. In vitro and in vivo studies demonstrated BBT-176's inhibitory potency against mutant EGFRs, achieving tumor regression in mouse models and showing early clinical efficacy in patients with EGFR 19Del/T790M/C797S mutation. These findings suggest BBT-176's potential as a fourth-generation EGFR inhibitor for treating NSCLC resistant to current EGFR TKIs.
Figure 1. The activity of BBT-176 against EGFR-mutated Ba/F3 cells and human cell lines was assessed. Cell growth inhibition curves and IC50 values were measured, indicating the potency of BBT-176 against EGFR mutants. (Lim SM, et al., 2023)
A: Ba/F3 cells were likely chosen for their cytokine-dependent growth and suitability for studying oncogenic kinase activity and drug resistance mechanisms associated with EGFR mutations such as DEL19 and C797S.
A: Stability was likely confirmed through methods such as immunoblotting, functional assays measuring downstream signaling, or cell viability assays in the absence of growth factors, with continuous selection pressure applied.
A: Characterization may involve analysis of EGFR phosphorylation, downstream signaling pathways, and functional implications in cell proliferation, survival, and response to EGFR inhibitors such as gefitinib or osimertinib.
A: Quality control likely included confirmation of EGFR-DEL19/C797S expression levels, validation of its kinase activity and drug sensitivity, assessment of off-target effects, and validation of phenotypic changes associated with EGFR modulation.
A: Comparative analysis with patient-derived samples or in vivo models helps validate the relevance of EGFR-DEL19/C797S expression in oncogenic signaling, tumor progression, and response to EGFR-targeted therapies, guiding the development of personalized treatment strategies for EGFR-mutant cancers, particularly those harboring resistance mutations like C797S.
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Unmatched stability! The Human EGFR-L858R/T790M/C797S Stable Cell Line in Ba/F3 cells ensures consistent expression of EGFR mutations, enabling reliable results in cancer drug resistance studies.
Empowering advanced investigations! With stable EGFR-L858R/T790M/C797S expression, I can explore mechanisms of resistance to EGFR inhibitors with confidence, driving progress in targeted therapy research.
Exceptional performance! This cell line surpasses expectations, providing a solid foundation for studying EGFR-L858R/T790M/C797S-targeted therapies and personalized medicine approaches in lung cancer.
Enhancing research efficiency! Its stable expression streamlines experimental workflows, facilitating streamlined data analysis and accelerating discoveries in drug resistance mechanisms.
An indispensable asset! The Human EGFR-L858R/T790M/C797S Stable Cell Line has revolutionized my research, offering valuable insights into EGFR-driven oncogenesis and potential therapeutic strategies for overcoming drug resistance.
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