Cat.No. : CSC-RO0656 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-RO0656 |
| Description | This cell line is engineered to stably overexpress exogenous human TPR-Met fusion sequence bearing D1228N mutation. |
| Target Gene | TPR-MET |
| 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 |
MET mutations, including exon 14 deletions and amplifications, are key drivers of non-small cell lung cancer (NSCLC), and targeted therapies against MET have shown clinical efficacy. However, resistance often develops, limiting the long-term success of these therapies. To address this challenge, the researchers examined secondary MET mutations that could arise from treatment with MET tyrosine kinase inhibitors (TKIs), including type I and type II inhibitors like capmatinib and merestinib. Using TPR-MET transformed Ba/F3 cells in mutagenesis assays, they identified distinct secondary mutant profiles associated with single-agent therapies. Notably, when capmatinib and merestinib were combined, no resistant clones emerged in vitro. In vivo, this combination significantly reduced tumor outgrowth compared to either drug alone, suggesting that the dual inhibition of MET could be an effective strategy to delay or overcome drug resistance.
Figure 1. The researchers used Ba/F3 cells expressing wild-type TPR-MET and mutagenized them with ENU to induce secondary MET mutations. These cells were exposed to various MET TKIs, and the emergence of drug-resistant clones was monitored. Resistant clones were isolated, expanded, and sequenced to identify mutations in the MET tyrosine kinase domain (TKD). Droplet digital PCR (ddPCR) assays were used to detect specific MET mutations like D1228N and G1163R. (Bahcall M, et al., 2022
Creative Biogene's Human TPR-MET-D1228N Stable Cell Line - Ba/F3 provides an ideal tool for studying MET mutations and the development of resistance mechanisms in cancer research. This cell line can be used for evaluating the efficacy of MET-targeted therapies, such as combination treatments with type I and II inhibitors.
A: Our Human TPR-Met-D1228N stable cell line exhibits a high degree of stability in long-term culture. We verified the stability of gene expression through PCR, Western blot and other methods, and conducted repeated tests at different time points and in different batches. The results showed that the cell line expression was stable with minimal fluctuations in gene expression levels.
A: The impact of the BaF3 cell line as a carrier on the Human TPR-Met-D1228N stable cell line is considered minimal. However, we recommend that customers pay attention to the special growth requirements of the BaF3 cell line during use to ensure optimal expression and stability of the cells. We provide detailed training guides to help clients overcome potential issues.
A: Yes, our BaF3 cell line has undergone a rigorous screening and optimization process. We use specific screening criteria, including antibiotic selection and gene expression levels, to ensure that the stability and expression levels of the cell lines meet our quality standards. We also performed long-term culture and expansion of the cell line to verify its stability and reproducibility under different conditions. We also use relevant cell function analysis methods, such as proliferation assays and functional assays, to ensure that the cell lines are performing as expected.
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While using this product, I experienced the professional quality of service provided by the supplier. They paid full attention to the stability of the cells to ensure consistent growth and stable gene expression during the culture process. The careful attention gives me great satisfaction, knowing that my research work can depend on this dependable stability.
The merchant handled the entire purchasing process in a very professional manner and provided timely guidance and support in the use of the product. I observed that the cells exhibited excellent stability during culture, which provided a solid foundation for my research. I'm so happy I chose them as my supplier.
Thanks to the excellent service provided by the supplier. They not only provided high-quality products but also gave helpful suggestions on cell growth and parameter evaluation. I observed that the cells showed good stability during the culture process, which made my research work progress smoothly. I would recommend them as a reliable supplier without hesitation.
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