Cat.No. : CSC-RO0306 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-RO0306 |
| Description | This cell line is engineered to stably overexpress human EGFR bearing D770del_ins_GY 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 |
| 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 |
Exon 20 insertion mutations in the Epidermal Growth Factor Receptor (EGFR) constitute approximately 10% of all EGFR mutations observed in lung cancers. Researchers utilized the Human EGFR-D770del_ins_GY Stable Cell Line to investigate EGFR exon 20 insertion mutations in lung cancer. This model helped identify variants, such as EGFR-D770>GY, responsive to approved 2nd generation EGFR-TKIs and EGFR exon 20 insertion mutant-active TKIs (poziotinib and mobocertinib). Clinical data supported preclinical findings, highlighting the efficacy of specific TKIs in treating EGFR exon 20 insertion mutants. These insights have implications for clinical care, genomic profiling, and clinical trial design.
Figure 1. The Human EGFR-D770del_ins_GY Stable Cell Line is utilized by researchers to probe EGFR-TKIs in Ba/F3 isogenic preclinical models of EGFR exon 20 insertion mutations. Sensitivity or resistance to treatment by various EGFR mutants, including EGFR-D770>GY, is assessed, revealing the therapeutic window of EGFR-TKIs. Dose-response proliferation assays with dacomitinib and afatinib further elucidate drug efficacy. (Kobayashi IS, et al., 2020)
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 ALK mutations, including the G1202R mutation.
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 ALK phosphorylation, downstream signaling pathways, and functional implications in cell proliferation, survival, and response to ALK inhibitors such as lorlatinib or brigatinib.
A: Quality control likely included confirmation of EML4-ALK-G1202R expression levels, validation of its kinase activity and drug sensitivity, assessment of off-target effects, and validation of phenotypic changes associated with ALK modulation.
A: Comparative analysis with patient-derived samples or in vivo models helps validate the relevance of EML4-ALK-G1202R expression in NSCLC progression, metastasis, and response to ALK-targeted therapies, guiding the development of personalized treatment strategies for patients with acquired resistance mutations.
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Unmatched reliability! The Human EGFR-D770del_ins_GY Stable Cell Line in Ba/F3 cells ensures stable expression of EGFR variants, providing consistent results in cancer drug sensitivity studies.
Enabling advanced exploration! With stable EGFR-D770del_ins_GY expression, I can investigate mechanisms of sensitivity to EGFR inhibitors with confidence, advancing personalized medicine research in lung cancer.
Remarkable performance! This cell line surpasses expectations, serving as a robust platform for studying EGFR-D770del_ins_GY-targeted therapies and precision treatment approaches in EGFR-mutant tumors.
Streamlining research workflows! Its stable expression simplifies experimental procedures, facilitating efficient data collection and analysis, and accelerating discoveries in drug sensitivity mechanisms.
An indispensable resource! The Human EGFR-D770del_ins_GY Stable Cell Line has revolutionized my research, offering valuable insights into EGFR-driven oncogenesis and potential therapeutic strategies for enhancing drug sensitivity in EGFR-mutated cancers.
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