Cat.No. : CSC-RO0138 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-RO0138 |
| Description | Ba/F3-EGFR-L858R/T790M cell line is a stably transfected cell line which expresses human epidermal growth factor receptor (EGFR) with L858R and T790M 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 |
Non-small cell lung cancer (NSCLC) with activating EGFR mutations was initially treated with first-generation EGFR tyrosine kinase inhibitors (EGFR-TKIs), but unfortunately, these drugs have limited efficacy, mostly due to the T790M mutation. Although osimertinib has been approved for the treatment of patients with T790M-positive NSCLC, most patients will develop C797S mutations and become ill again. Therefore, more new treatment strategies for T790M mutation-positive NSCLC are urgently needed. It was hypothesized that wighteone, a natural compound isolated from plant derivatives, would have antitumor effects against NSCLC with T790M mutation. In this study, a Ba/F3 cell line carrying EGFR L858R/T790M mutations (Ba/F3 EGFR L858R/T790M cell line) was established, and the effects and mechanisms of wighteone were investigated using this cell line and a human NSCLC cell line carrying EGFR L858R/T790M mutations (NCI-H1975). The results showed that wighteone inhibited cell proliferation, suppressed EGFR signaling pathway, caused cell cycle redistribution and induced cell apoptosis. The study showed that wighteone may provide a new potential treatment strategy for NSCLC patients carrying T790M mutation.
To evaluate the effect of leukotoxin on proliferation, the researchers first tested a range of concentrations of leukotoxin in Ba/F3 cells, Ba/F3 EGFR L858R/T790M cells, and human NSCLC cells NCI-H1975. Compared with Ba/F3 cells, leukotoxin (Figure 1A) had a stronger inhibitory effect in Ba/F3 EGFR L858R/T790M cells and NCI-H1975 cells (Figure 1B-D). Then, colony formation assays were further performed to determine the antiproliferative potential of leukotoxin. As shown in Figure 1E, F, wighteone significantly inhibited the clonogenic potential of NCI-H1975 cells in a concentration-dependent manner. These results proved the effect of wighteone on cell proliferation against EGFR L858R/T790M.
Figure 1. Wighteone effectively inhibits EGFR L858R/T790M. (A) Chemical structure of Wighteone. The inhibitory effect of Wighteone was evaluated in Ba/F3 (B), Ba/F3 EGFR L858R/T790M (C), and NCI-H1975 (D) cells. (E) NCI-H1975 cells were treated with different concentrations of Wighteone, and the colony formation efficiency was observed by colony formation assay. (F) Quantitative results of clonogenic effects were analyzed. (Sun P, et al., 2021)
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We have been using the human EGFR-L858R/T790M stable cell line - Ba/F3 for cancer drug screening projects, and it has consistently provided reliable and reproducible results.
The Human EGFR-L858R/T790M Stable Cell Line-Ba/F3 has proven to be extremely valuable in our drug resistance studies. Its relevance to NSCLC research has helped us gain a deeper understanding of resistance mechanisms and supported the development of targeted therapies.
Since incorporating the Human EGFR-L858R/T790M Stable Cell Line-Ba/F3 into our research pipeline, we have been able to accelerate the drug development process. We highly recommend this product to any laboratory focused on oncology research.
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