Cat.No. : CSC-RO1117 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-RO1117 |
| Description | This cell line is engineered to stably overexpress human MET in Ba/F3 cells. |
| Target Gene | 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 |
Hepatocyte growth factor (HGF) activates the c-MET receptor, which is crucial for tissue regeneration and cellular processes like migration and proliferation. However, the clinical application of HGF is limited due to its instability and poor recombinant expression. The researchers engineered a more stable and expressible HGF fragment, eNK1, and developed a dimerized version of eNK1 that showed significantly improved c-MET activation and cellular responses compared to the monomer. In this study, the researchers used BaF3-MET cells, a murine cell line transfected with human c-MET receptors, to analyze the binding affinity and activity of the eNK1 monomer and dimer. They found that the eNK1 dimer had a much lower IC50, indicating stronger binding affinity. This research highlights the potential of Creative Biogene's Human MET Stable Cell Line - BaF3 as a tool for similar studies on c-MET receptor activation and its implications for regenerative medicine
Figure 1. The researchers treated HUVEC cells with HGF, eNK1 monomer, or eNK1 dimer at various concentrations to assess c-MET activation. They used Western blot analysis to detect phosphorylated c-MET, Akt, and Erk and employed the PathHunter system to quantify c-MET activation through chemiluminescence. BaF3-MET cells were used for binding assays to measure affinity. (Hassannia H, et al., 2022)
A: Our Human MET stable cell line has undergone rigorous screening and verification to ensure stable expression of the MET gene. We verified the stability of the MET gene through PCR, Western blot and other methods, and conducted repeated tests at different time points and in different batches. Cells express MET under appropriate culture conditions and maintain stable expression levels.
A: Our Human MET stable cell line exhibits normal growth and proliferation properties on a BaF3 background. We provide detailed culture guidelines to help customers overcome the potential unique growth requirements of the BaF3 cell line. Customers are advised to pay close attention to cell growth during use and follow our culture recommendations.
A: Yes, our Human MET stable cell lines have passed strict quality control inspections. We verify cell lines according to international and internal standards, including stability of gene expression, cell morphology, contaminant detection, etc. We ensure that each batch of cell lines meets our quality standards to ensure our customers receive a high-quality product.
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This human MET stable cell line expresses very reliably in BaF3 cells. In my experiments, I observed that these cells stably expressed MET receptor tyrosine kinase and responded significantly to HGF binding. This process is of great significance to cell survival, embryonic development, migration and invasion. This cell line provides a reliable platform for studying the activity of the MET signaling pathway and the pathogenesis of related diseases.
Stable cell lines expressing human MET in BaF3 cells showed outstanding performance. It was observed that after these cells were stimulated by HGF, the MET receptor dimerized and was activated, triggering a series of signal transduction reactions. This provides an ideal model for us to study the role of the MET signaling pathway in biological processes such as cell migration and invasion, and helps us better understand the pathogenesis of related diseases.
The application of the human MET stable cell line in BaF3 cells has been very satisfactory. Through my experimental observations, these cells can stably express MET receptors and respond significantly to HGF binding. This provides an important tool for us to further study the role of the MET signaling pathway in malignant tumors such as hepatocellular carcinoma and head and neck tumors, and helps us find therapeutic targets for related diseases.
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