Cat.No. : CSC-RO0708 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-RO0708 |
| Description | Ba/F3-KIF5B-RET-G810S cell line is engineered to stably overexpress human KIF5B-RET fusion protein bearing G810S mutation in RET part. |
| Target Gene | KIF5B-RET |
| 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 |
Alterations in the RET gene, a receptor tyrosine kinase, are frequently observed in thyroid and lung cancers, where they contribute to tumor growth. In recent years, RET inhibitors have been used in clinical settings to treat thyroid cancer and are being explored in clinical trials for RET fusion-positive non-small cell lung cancer (NSCLC). However, the role of mutations in the RET kinase domain, which may confer resistance to these therapies, remains an area of ongoing research. The researchers used BaF3/KIF5B-RET cells to investigate how mutations in the RET kinase domain impact the effectiveness of multiple RET-targeted tyrosine kinase inhibitors (TKIs), including cabozantinib, lenvatinib, vandetanib, and nintedanib. By analyzing fourteen mutations, they identified several variants, such as G810S, that exhibited pan-resistance to all four TKIs. These findings highlight the importance of understanding RET mutation profiles to select the most effective treatment options.
Figure 1. The researchers treated BaF3/KIF5B-RET cells and its RET kinase domain mutants with cabozantinib, lenvatinib, or vandetanib at various concentrations. (Liu X, et al., 2018)
Creative Biogene's Human KIF5B-RET-G810S Stable Cell Line - Ba/F3 offers a valuable model for studying RET kinase mutations and their impact on drug resistance, helping advance personalized cancer therapies.
A: Our Human KIF5B-RET-G810S 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 Ba/F3 cell line as a carrier on the Human KIF5B-RET-G810S stable cell line is considered minimal. However, we recommend that customers pay attention to the special growth requirements of the Ba/F3 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 Ba/F3 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. In addition, relevant functional analyzes were performed on the cell lines to ensure that they performed as expected.
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This human KIF5B-RET-G810S stable cell line expressed excellently in Ba/F3 cells, and I noticed stable cell growth and consistent gene expression. This stability allows us to conduct cell experiments with greater confidence without worrying about variation or unstable expression. The cell growth was also satisfactory and in line with our expectations.
Expression in Ba/F3 cells was satisfactory for the human KIF5B-RET-G810S stable cell line from this supplier. Through precise manipulation of genetic engineering technology, cells stably express the RET fusion protein and maintain the G810S mutation. During the cell culture process, we observed that the cells grew well and gene expression remained consistent. This allows us to obtain reliable results in experiments and conduct in-depth studies of the biological properties of cells.
I would like to give a positive word about this human KIF5B-RET-G810S stable cell line. This cell line showed satisfactory stability and consistent gene expression in Ba/F3 cells. Through the precise design of genetic engineering technology, cells stably express the KIF5B-RET fusion protein while maintaining the G810S mutation. During the cell culture process, we observed that the cells grew well and gene expression was stable, which provided a reliable basis for our research. In addition, Ba/F3 cells are relatively easy to culture and genetically modify, allowing us to conduct experiments efficiently and explore different signaling pathways and biological processes.
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