Cat.No. : CSC-RO0831 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-RO0831 |
| Description | The Ba/F3-EPOR-JAK2-V617F cell line is engineered to stably overexpress exogenous human EPOR-JAK2 bearing V617F amino acid mutation in the JAK2 part. |
| Target Gene | EPOR-JAK2 |
| 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 |
A common mutation, JAK2 V617F, plays a pivotal role in these conditions, often leading to resistance against conventional type I JAK inhibitors. In recent investigations, the efficacy of a type II JAK inhibitor, CHZ868, was evaluated in JAK inhibitor-persistent MPN cells. The study found that cells harboring the JAK2 V617F mutation demonstrated significant sensitivity to CHZ868, achieving a marked reduction in mutant allele burden—an outcome not seen with standard treatments. These insights reveal the potential for type II JAK inhibitors to serve as effective therapeutic alternatives for patients with JAK2-related MPNs.
Figure 1. The researchers employed the Human EPOR-JAK2-V617F Stable Cell Line - Ba/F3 to evaluate the proliferation and apoptotic responses to various JAK inhibitors. Their goal was to elucidate the differential effects of CHZ868 compared to standard treatments, informing future therapeutic strategies for MPNs. (Meyer SC, et al., 2015)
The Human EPOR-JAK2-V617F Stable Cell Line - Ba/F3 developed by Creative Biogene offers a valuable resource for researchers exploring the mechanisms of JAK2 signaling and testing new therapeutics. This cell line mirrors the disease's biological features, making it ideal for drug efficacy studies and molecular investigations.
A: The Ba/F3 cell line expressing the human EPOR and the JAK2-V617F mutation provides a controlled system to investigate the signaling cascades triggered by erythropoietin binding to EPOR. Researchers can use this cell line to study the activation of downstream effectors such as STAT5, which is critical for erythroid cell differentiation and proliferation. By introducing specific inhibitors or activators of the JAK2-V617F pathway, the cell line can be used to dissect the molecular mechanisms underlying the enhanced signaling and the potential therapeutic targets for diseases like polycythemia vera.
A: For high-throughput screening, it is essential to establish a robust assay protocol that measures the activity of the EPOR/JAK2 signaling axis. This may involve the use of specific reporter genes under the control of EPOR-responsive promoters or the measurement of phosphorylation events using flow cytometry or Western blotting. Researchers must ensure that the cell line is maintained under optimal conditions, and that the screening process is standardized to minimize variability. Additionally, it is crucial to validate the hits from the screen using secondary assays to confirm their specificity and efficacy.
A: By utilizing the Ba/F3 cell line with the EPOR-JAK2-V617F system, researchers can model the hyperactive signaling characteristic of myeloproliferative neoplasms. Through the introduction of mutations known to be associated with these disorders, the cell line can be used to study the molecular pathways that drive disease progression. Moreover, this system can be employed to test the efficacy of targeted therapies, such as JAK2 inhibitors, which are currently used in the treatment of myeloproliferative neoplasms.
A: The Ba/F3 cell line is known for its ability to be genetically manipulated and its reliance on cytokines, like erythropoietin, for growth, making it an excellent model for studying cytokine signaling pathways. The stable expression of the EPOR and the JAK2-V617F mutation allows for consistent and controlled experiments, which is particularly beneficial for functional studies. This system provides a more physiologically relevant context compared to transient transfection systems, as it allows for the study of long-term effects and the establishment of signaling pathways.
A: The Ba/F3 cell line with the integrated EPOR-JAK2-V617F system offers a unique platform to explore the interplay between erythropoietin signaling and other pathways that are critical for hematopoiesis. Researchers can use this cell line to investigate how EPOR activation influences other signaling molecules, such as those involved in cell survival, differentiation, and apoptosis. This can lead to a better understanding of the complex regulatory networks in hematopoietic cells and may reveal novel therapeutic targets for blood disorders.
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The Human EPOR-JAK2-V617F Stable Cell Line - Ba/F3 precisely models the JAK2 V617F mutation, common in myeloproliferative disorders. This precision allows for exact replication of the mutation's effects on cell signaling and proliferation, crucial for studying its specific impact on cellular processes.
This cell line ensures consistent expression of the JAK2 V617F mutation alongside the EPOR receptor, providing stable experimental conditions. The Human EPOR-JAK2-V617F Stable Cell Line - Ba/F3 is essential for obtaining reproducible results in research focusing on the interactions and pathways influenced by this mutation.
Leveraging the robust growth characteristics of the Ba/F3 cells, the Human EPOR-JAK2-V617F Stable Cell Line - Ba/F3 supports extensive experimental use without loss of phenotype. This robust growth is advantageous for long-term studies and ensures the durability of the cell line over numerous passages.
With its stable genetic background and consistent protein expression, the Human EPOR-JAK2-V617F Stable Cell Line - Ba/F3 streamlines the process of data collection, reducing variability and enhancing the accuracy of experimental results. This streamlined data collection is crucial for drawing reliable conclusions in signal pathway research.
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