Cat.No. : CSC-RO0135 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-RO0135 |
| Description | Ba/F3-EGFR-H773_V774insNPH cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with H773_V774insNPH 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 |
In non-small-cell lung cancer (NSCLC), uncommon epidermal growth factor receptor (EGFR) mutations account for 10%–20% of all EGFR mutations. Mutations often impair the efficacy of traditional medicines like osimertinib and afatinib. Thus, novel EGFR-tyrosine kinase inhibitors are needed. Aumolertinib, a third-generation EGFR-TKI, may cure advanced NSCLC, however its effectiveness against uncommon mutations is uncertain. In this work, engineered Ba/F3 cells and patient-derived cells with diverse rare EGFR mutations were used to explore in vitro anticancer efficacy of aumolertinib. Results indicated that aumolertinib effectively inhibited the viability of cell lines harboring these mutations, outperforming its effects on wild-type EGFR cells. Additionally, aumolertinib demonstrated significant tumor growth inhibition in mouse models with specific uncommon mutations, suggesting its potential as a viable treatment option.
Figure 1. The researchers evaluated the antitumor activity of aumolertinib using a patient-derived xenograft (PDX) model featuring the EGFR H773-V774insNPH mutation. (Shi C, et al., 2016)
Creative Biogene's Human EGFR-H773_V774insNPH Stable Cell Line - Ba/F3 provides a valuable tool for further exploring the therapeutic implications of aumolertinib and similar agents. Our cell line may facilitate rigorous testing of new compounds, ensuring that emerging therapies can be evaluated for effectiveness against hard-to-treat NSCLC variants.
A: The EGFR-H773_V774insNPH represents a novel insertion mutation identified in the epidermal growth factor receptor (EGFR) gene. This mutation, located in exon 20, involves the insertion of the sequence "NPH" (asparagine-proline-histidine) between amino acids 773 and 774 of the EGFR protein. This insertion mutation can lead to aberrant EGFR signaling and may contribute to the development of resistance to EGFR-targeted therapies in certain cancers, particularly non-small cell lung cancer (NSCLC).
A: The EGFR-H773_V774insNPH mutation affects EGFR function by altering the structure and activity of the kinase domain. This insertion can lead to the formation of a novel activation loop, which enhances the kinase activity of the EGFR protein. As a result, the mutated EGFR can drive uncontrolled cell growth and survival, contributing to the development of cancer.
A: Targeting the EGFR-H773_V774insNPH mutation has therapeutic implications for the treatment of cancers that harbor this specific mutation. Inhibitors that can specifically target this mutation, such as osimertinib (Tagrisso), may be effective in treating patients with NSCLC and acquired resistance to first-line EGFR inhibitors. These drugs can lead to tumor regression and improve patient outcomes.
A: The EGFR-H773_V774insNPH mutation is detected through molecular diagnostic techniques such as next-generation sequencing (NGS), which can identify the specific insertion mutation in the EGFR gene. This allows for accurate diagnosis and prognosis of patients with NSCLC and acquired resistance to EGFR inhibitors. Reverse transcription polymerase chain reaction (RT-PCR) and other molecular diagnostic methods can also be used to detect this mutation.
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
The Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3 is a result of meticulous product development, providing researchers with a valuable tool to study the effects of specific EGFR mutations on cell behavior and response to potential therapeutics.
This cell line demonstrates high knockout efficiency, ensuring that the desired mutation is effectively incorporated into the EGFR gene. This efficiency allows researchers to confidently study the biological consequences of this specific mutation.
The Human EGFR-H773_V774insNPH Stable Cell Line-Ba/F3 exhibits high genetic stability, maintaining the mutation over multiple cell divisions. This stability is crucial for conducting long-term experiments and ensures consistent and reliable results.
The cell line shows robust growth and maintains a normal morphology, which is essential for accurate interpretation of experimental results. The consistent cell growth allows researchers to perform experiments with high reproducibility and reliability.
Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.
Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Copyright © Creative Biogene. All rights reserved.
