Human EGFR-A763_Y764insFQEA Stable Cell Line - BaF3

Name: Human EGFR-A763_Y764insFQEA Stable Cell Line - BaF3
SKU: CSC-RO0302
Rating: 4.0 (3 reviews)

Cat.No. : CSC-RO0302 Host Cell: Ba/F3

Size: >1x10^6 frozen cells/vial, 1 mL Stability: Stable in culture over a minimum of 10 passages

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Cat. No.	CSC-RO0302
Description	This cell line is engineered to stably overexpress human EGFR bearing A763_Y764insFQEA 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

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EGFR-A763_Y764insFQEA is a mutation found in the epidermal growth factor receptor (EGFR) gene. This is a specific type of insertional mutation, which means that extra DNA base pairs (in this case, FQEA) are added to the coding sequence at positions 763 and 764.  In normal circumstances, the EGFR gene produces a protein that helps cells, particularly skin cells, to grow and divide. However, the A763_Y764insFQEA mutation causes the gene to function abnormally, which often leads to uncontrolled cell growth and the development of potential cancer, most commonly lung cancer. This type of mutation is typically discovered through DNA sequencing or genomic testing and, although rare, may be important in personalized cancer treatment, particularly in predicting the likelihood of response or resistance to certain targeted therapies. 

The human EGFR-A763_Y764insFQEA stable cell line - BaF3 is a powerful research tool designed to study specific forms of EGFR involved in tumor proliferation and resistance to EGFR-targeted therapies. The BaF3 cell line is a mouse pro-B cell line that lacks expression of functional EGFR, making it a suitable background for expressing mutant forms of EGFR. The human EGFR-A763_Y764insFQEA stable cell line could help researchers develop and test new treatments for cancers caused by this specific EGFR mutation. This cell line is ready-to-use and guaranteed to provide consistent research results.

EGFR exon 20 insertion mutations account for 5% to 10% of EGFR-mutant NSCLC. CLN-081 (formerly known as TAS6417) is a novel covalent EGFR tyrosine kinase inhibitor that has demonstrated pan-mutation selective efficacy in the clinical setting, including exon 20 insertions. However, some patients may not respond to CLN-081, while others may develop resistance to CLN-081 over time. Studies here demonstrate that all CLN-081-resistant clones harbor the EGFR C797S mutation. Ba/F3 cells with C797S (Ba/F3-C797S) are resistant to EGFR tyrosine kinase inhibitors targeting EGFR exon 20 insertion mutations, including CLN-081. Pimitespib is a selective inhibitor of heat shock protein 90 that induces apoptosis in Ba/F3-C797S cells in vitro and inhibits the growth of Ba/F3-C797S tumors in vivo. Ba/F3 cells with A763_Y764insFQEA-C797S remain sensitive to erlotinib.

Figure 1. Cells containing EGFR A763_Y764insFQEA (insFQEA) remain sensitive to erlotinib. (A) Cell viability assays were performed after treatment of Ba/F3-insFQEA (EGFR-A763_Y764insFQEA Cell Line - BaF3) or Ba/F3-insFQEA-C797S cells with different concentrations of EGFR TKI for 48 hours. (B) IC50 ratios of the indicated drugs (Ba/F3-insFQEA-C797S relative to Ba/F3-insFQEA) according to the cell viability assay described in (A). (C) Western blots showing EGFR and downstream signaling factors after erlotinib treatment of Ba/F3-insFQEA-C797S cells. (D) Apoptosis assays were performed using Annexin V staining in Ba/F3-insFQEA-C797S cells treated with CLN-081, erlotinib, or vehicle control for 24 hours. (E) Effects of erlotinib or vehicle on a mouse xenograft tumor model containing Ba/F3-insFQEA-C797S cells. (F) Tumor weights of mice analyzed in (E). (Kagawa Y, et al. 2023)

The human EGFR-A763_Y764insFQEA stable cell line - BaF3 is widely used in biomedical research and pharmaceutical industries for a variety of applications.

Drug development: This cell line serves as a stable model system for screening and evaluating potential drugs targeting the EGFR-A763_Y764insFQEA mutation. It allows researchers to study the effects of these drugs on cell viability, proliferation and apoptosis.

Molecular biology research: It helps scientists understand the underlying biological mechanisms associated with the EGFR-A763_Y764insFQEA mutant. This cell line helps study the function and behavior of this mutant protein at the molecular level.

Cancer research: Given the critical role of EGFR mutations in various types of cancer, especially lung cancer, this cell line is widely used in oncology research to study cancer pathology, tumorigenesis processes, and drug resistance issues.

Protein expression studies: This cell line is valuable for examining the effects of mutant protein expression on cell phenotypes, signal transduction pathways, and other cellular processes.

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Customer Reviews

Unmatched analytical value

The analytical value of the Human EGFR-A763_Y764insFQEA Stable Cell Line - BaF3 is unmatched. It has greatly enhanced our ability to study the specific insertion mutation and its implications in cancer biology. The data produced has been instrumental in advancing our research objectives. Highly recommended for anyone focused on EGFR-related studies.

United States

02/05/2022

Superior Performance in In Vitro Studies

The Human EGFR-A763_Y764insFQEA Stable Cell Line - BaF3 has demonstrated superior performance in our assays, enabling us to generate high-quality data quickly and efficiently. It has become a cornerstone of our experimental setup.

United States

03/14/2023

High-quality and reliable

The cells exhibit robust growth and the genetic stability is spot-on, which gives us confidence in our data. It has greatly facilitated our research in understanding EGFR-driven oncogenic pathways.

United States

03/22/2024

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Human EGFR-A763_Y764insFQEA Stable Cell Line - BaF3

Cell Line Information

Cell Culture Information

Safety and Packaging

Background

Case Study

Applications

Publications

Q & A

Customer Reviews

Unmatched analytical value

Superior Performance in In Vitro Studies

High-quality and reliable

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