Cat.No. : CSC-RO0394 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-RO0394 |
| Description | This cell line is engineered to stably overexpress exogenous human KIF5B-RET fusion protein. |
| 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 |
Both thyroid and lung cancers include changes in the tyrosine kinase enzyme RET. Using the Human KIF5B-RET Stable Cell Line - BaF3, the scientists looked into how different tyrosine kinase inhibitors (TKIs) affected the sensitivity of RET kinase domain mutations. They found and examined mutations that conferred resistance to TKIs such as cabozantinib, lenvatinib, vandetanib, and nintedanib using RET kinase-dependent BaF3/KIF5B-RET cells. Additionally, they investigated the TKIs' ability to affect the RET (M918T) mutation, which is frequently detected in aggressive multiple endocrine neoplasia type 2B. The research demonstrated that particular mutations resulted in either pan resistance or selective resistance to the medications, offering a guide for choosing the right TKIs for RET kinase domain mutants.
Figure 1. To evaluate the dose-dependent suppression of different RET kinase domain mutations using cabozantinib, lenvatinib, and vandetanib, the researchers created a Human KIF5B-RET Stable Cell Line - BaF3. They also used immunoblotting to analyze cell survival and protein expression. (Liu X, et al., 2018)
A: Our Human KIF5B-RET stable cell line exhibits a high degree of stability after multiple cultures and expansions. We verified the stability of the cell lines through a variety of methods, including long-term culture, drug screening, and PCR. Under appropriate culture conditions, cell line expression is stable and the original expression level of the gene is maintained. In addition, we provide detailed culture guidelines to ensure customers can maintain stable expression of their cell lines.
A: We performed a comprehensive assessment of cell proliferation and metabolic activity, and the results showed that this cell line exhibited normal cell proliferation and metabolic activity under normal culture conditions. We recommend that customers undergo a period of adaptation culture after receiving the cell lines to ensure optimal growth and activity of the cells.
A: Yes, our Human KIF5B-RET stable cell line has 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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The stability and professionalism of this product are impressive. During use, I noticed stable cell growth and consistent gene expression. This is very important for my experiments because it ensures that the data I get are accurate and reliable. In addition, the service of the product supplier is also very considerate. They answered my questions in a timely manner and ensured my understanding and use of the product.
For my research, this human KIF5B-RET stable cell line provides a valuable tool. Not only does it provide stability of gene expression, it also models the pathogenesis of lung adenocarcinoma, allowing me to better understand this disease. In addition, the platform provided by the supplier is also very convenient, and I can easily obtain the product information I need and make purchases.
The human KIF5B-RET stable cell line performed excellently in my experiments. I observed stable cell growth and consistent gene expression. This is very important for my research because it ensures the reliability of the data I get. The service provided by the supplier is also very good. They answered my questions in a timely manner and made me feel more confident about using the product.
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