Cat.No. : CSC-RO0143 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-RO0143 |
| Description | Ba/F3-EGFR-T790M/L861Q cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with T790M and L861Q 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 |
Especially T790M and L861Q, EGFR mutations are often linked to non-small cell lung cancer (NSCLC) and cause resistance to first- and second-generation EGFR inhibitors. Examining SH-1028, a unique third-generation EGFR TKI, the researchers assessed its efficacy against both EGFR-sensitive and -resistant mutations including T790M and L861Q. With considerably stronger efficacy against L858R/T790M mutants than wild-type EGFR, SH-1028 showed strong suppression of mutant EGFR kinases in enzymatic tests. Without inducing notable weight loss, in vivo studies revealed that SH-1028 drastically slowed tumor development in xenograft models with EGFR mutations—including the resistant T790M mutant.
Figure 1. The researchers evaluated the preliminary efficacy of SH-1028 by conducting a kinase inhibition assay against various EGFR mutations, including WT, L858R, d746-750, L861Q, T790M/L858R, and T790M/d746-750. (Han L, et al., 2021)
Creative Biogene's Human EGFR-T790M/L861Q Stable Cell Line-Ba/F3 may be used to explore EGFR inhibitors, especially new medicines targeting these mutations. This cell line may help researchers study drug resistance and improve EGFR-targeted medicines.
A: The EGFR-T790M/L861Q gene represents a specific mutation in the EGFR (epidermal growth factor receptor) gene, which is associated with resistance to EGFR-targeted therapies in non-small cell lung cancer (NSCLC) and other cancers. The T790M and L861Q mutations occur in the tyrosine kinase domain of the EGFR protein, leading to drug resistance by activating alternative signaling pathways.
A: The T790M and L861Q mutations in the EGFR protein affect its function by altering the kinase domain. These changes can lead to increased kinase activity, which in turn promotes uncontrolled cell growth and survival. The T790M mutation specifically confers resistance to first-line EGFR inhibitors like gefitinib and erlotinib.
A: Targeting the EGFR-T790M/L861Q mutation has significant therapeutic implications. Inhibitors that can overcome the resistance caused by these mutations, such as osimertinib (Tagrisso), are used to treat patients with NSCLC and this specific mutation. These drugs can lead to tumor regression and improve patient outcomes in the second-line setting.
A: The EGFR-T790M/L861Q mutation is detected through molecular diagnostic techniques such as reverse transcription polymerase chain reaction (RT-PCR), fluorescence in situ hybridization (FISH), and next-generation sequencing (NGS). These methods can identify the specific T790M and L861Q mutations, allowing for accurate diagnosis and treatment planning for patients with NSCLC and acquired resistance to EGFR inhibitors.
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The Human EGFR-T790M/L861Q Stable Cell Line-Ba/F3 is a reliable tool for studying EGFR mutation-driven tumors, providing consistent and valuable insights into cancer biology.
The cell line comes with comprehensive documentation and accessible technical support, facilitating successful experimentation and troubleshooting for researchers.
The Human EGFR-T790M/L861Q Stable Cell Line-Ba/F3 is ethically sourced and adheres to relevant regulations, ensuring responsible and ethical research practices.
The cell line is designed for safe handling and cultivation in the laboratory, reducing the risk of contamination and genetic alterations, and promoting a secure research environment.
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