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  6. Human EGFR-D770del_ins_GY Stable Cell Line - Ba/F3

Human EGFR-D770del_ins_GY Stable Cell Line - Ba/F3

For research use only. Not intended for any clinical use.

Cat. No. :   CSC-RO0306

Host Cell :   Ba/F3 Size :   >1x106 frozen cells/vial

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Cell Line Information

Cell Culture Information

Safety and Packaging

Gene Information

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)
Applications Drug screening and biological assays
Biosafety Level 2
Size >1x10^6 frozen cells/vial, 1 mL
Stability Stable in culture over a minimum of 10 passages
Quality Control Negative for bacteria, yeast, fungi and mycoplasma.
Growth Conditions 37 °C, 5% CO2
Storage Liquid nitrogen
Shipping Dry ice
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.
Freeze Medium Frozen with 70% medium, 20% FBS, 10% DMSO
Growth Properties Suspension, round
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
Gene Name EGFR epidermal growth factor receptor [ Homo sapiens ]
Gene Symbol EGFR
Synonyms ERBB; HER1; mENA; ERBB1; PIG61
Gene Description epidermal growth factor receptor (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian)
Gene ID 1956
Uni Prot ID P00533
m RNA Refseq NM_005228.3
Protein Refseq NP_005219.2
Chromosome Location 7p12
Function ATP binding; MAP kinase kinase kinase activity; actin filament binding; double-stranded DNA binding; enzyme binding; epidermal growth factor-activated receptor activity; epidermal growth factor-activated receptor activity; identical protein binding; contributes_to nitric-oxide synthase regulator activity; protein binding; protein heterodimerization activity; protein phosphatase binding; protein tyrosine kinase activity; protein tyrosine kinase activity; protein tyrosine kinase activity; receptor signaling protein tyrosine kinase activity; transmembrane receptor protein tyrosine kinase activity; transmembrane signaling receptor activity;
Pathway Adaptive Immune System, organism-specific biosystem; Adherens junction, organism-specific biosystem; Adherens junction, conserved biosystem; Alpha6-Beta4 Integrin Signaling Pathway, organism-specific biosystem; Androgen Receptor Signaling Pathway, organism-specific biosystem; Arf6 signaling events, organism-specific biosystem; Axon guidance, organism-specific biosystem;
MIM 131550
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The epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein belonging to the ErbB family of receptor tyrosine kinases (RTKs), pivotal in regulating cell growth, proliferation, and survival. The discovery of EGFR and its variants, such as D770 and G1202R mutations, has significantly impacted cancer research, particularly in understanding resistance mechanisms to EGFR-targeted therapies. These mutations alter the receptor's structure, leading to dysregulated signaling pathways implicated in tumorigenesis.

The BaF3 cell line, originally derived from murine pro-B cells, has been extensively utilized as a model system for studying EGFR signaling due to its ease of manipulation and growth factor dependency. The establishment of stable BaF3 cell lines expressing human EGFR-D770del_ins_GY mutation represents a crucial milestone in elucidating the functional consequences of this mutation and its role in cancer progression. This cellular model facilitates in-depth investigations into the molecular mechanisms underlying EGFR-driven malignancies, offering invaluable insights into potential therapeutic strategies.

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.

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.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)

1. Drug Screening: Employed to assess efficacy of EGFR inhibitors against D770del_ins_GY mutation in BaF3 cells.

2. Mechanistic Studies: Investigating signaling pathways activated by EGFR-D770del_ins_GY to elucidate its oncogenic mechanisms.

3. Therapeutic Development: Assessing novel targeted therapies specifically tailored for EGFR-D770del_ins_GY-positive cancers.

4. Resistance Mechanisms: Studying how cancer cells develop resistance to EGFR inhibitors targeting D770del_ins_GY mutation.

5. Preclinical Models: Utilized for in vivo studies to evaluate the efficacy and safety of potential therapeutic agents targeting EGFR-D770del_ins_GY mutation.

Customer Q&As
What influenced the selection of Ba/F3 cells for establishing the stable EML4-ALK-G1202R cell line?

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.

How was the stability and expression level of EML4-ALK-G1202R verified and maintained in this Ba/F3 stable cell line?

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.

Can you describe the characterization of EML4-ALK-G1202R expression in the Ba/F3 stable cell line, including its kinase activity and sensitivity to ALK inhibitors?

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.

What quality control measures were implemented during the generation of this stable cell line?

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.

How do the observed functional properties of EML4-ALK-G1202R in this stable cell line relate to its relevance in studying acquired resistance to ALK-targeted therapies in non-small cell lung cancer (NSCLC)?

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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Customer Reviews
Reliable EGFR-D770del_ins_GY Stable Cell Line

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.

United States

Empowering sensitivity mechanism exploration

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.

Germany

Remarkable EGFR-D770del_ins_GY performance

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.

French

Streamlined research workflows

Streamlining research workflows! Its stable expression simplifies experimental procedures, facilitating efficient data collection and analysis, and accelerating discoveries in drug sensitivity mechanisms.

United States

Invaluable resource for drug sensitivity studies

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.

Germany

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