Cat.No. : CSC-RO0241 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-RO0241 |
| Description | This cell line stably expresses human FLT3 gene bearing ITD duplicate and F691L amino acid double mutations. |
| Target Gene | FLT3 |
| 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. |
| Gene Name | FLT3 fms-related tyrosine kinase 3 [ Homo sapiens ] |
| Gene Symbol | FLT3 |
| Synonyms | FLK2; STK1; CD135; FLK-2 |
| Gene Description | fms-related tyrosine kinase 3 |
| Gene ID | 2322 |
| UniProt ID | P36888 |
| mRNA Refseq | NM_004119.2 |
| Protein Refseq | NP_004110.2 |
| Chromosome Location | 13q12 |
| Function | ATP binding; cytokine receptor activity; phosphatidylinositol 3-kinase binding; protein binding; protein homodimerization activity; transmembrane receptor protein tyrosine kinase activity; vascular endothelial growth factor-activated receptor activity; |
| Pathway | Acute myeloid leukemia, organism-specific biosystem; Acute myeloid leukemia, conserved biosystem; Cytokine-cytokine receptor interaction, organism-specific biosystem; Cytokine-cytokine receptor interaction, conserved biosystem; Hematopoietic cell lineage, organism-specific biosystem; Hematopoietic cell lineage, conserved biosystem; Pathways in cancer, organism-specific biosystem; |
| MIM | 136351 |
| 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 |
FLT3-ITD mutations are frequent in acute myeloid leukemia (AML) and have a poor prognosis. TKIs have improved, yet relapse and treatment resistance persist. In this work, researchers tested ningetinib, a new TKI, against FLT3-mutated AML cells. They performed cell proliferation assays using AML cell lines and Ba/F3 cells expressing different FLT3 mutations to evaluate the antileukemic potential of ningetinib. The study also included molecular docking and CETSA assays to validate ningetinib's interaction with FLT3 and its inhibition of key signaling pathways like STAT5, AKT, and ERK.
Figure 1. The researchers utilized a leukemia mouse model induced by Ba/F3-FLT3-ITD-F691L cells to evaluate ningetinib's efficacy in vivo. Flow cytometry was employed to measure GFP-positive cells in peripheral blood, bone marrow, and spleen, while survival curves and organ weights were analyzed to assess the therapeutic effects of ningetinib compared to other TKIs. (Hu C, et al., 2024)
Creative Biogene's Human FLT3-ITD-F691L Stable Cell Line-Ba/F3 can be utilized to study similar mutations and evaluate drug efficacy. Our stable cell line provides a reliable model for investigating the mechanisms of drug resistance and for screening potential therapies targeting FLT3 mutations. This cell line is valuable for researchers developing new treatments for AML, particularly in the context of overcoming resistance to existing TKIs like gilteritinib or quizartinib.
A: The FLT3-ITD-F691L gene represents a specific mutation in the FLT3 (fms-like tyrosine kinase 3) gene, which is found in some cases of acute myeloid leukemia (AML). The ITD (internal tandem duplication) mutation leads to the production of an aberrant FLT3 protein with increased tyrosine kinase activity. The F691L mutation is a further alteration within the kinase domain that enhances this activity, driving leukemogenesis.
A: The FLT3-ITD-F691L mutation contributes to leukemogenesis by activating the FLT3 signaling pathway in an uncontrolled manner. The mutated FLT3 protein has increased tyrosine kinase activity, leading to unregulated cell growth, survival, and differentiation. This dysregulation can disrupt normal hematopoiesis and promote the development of AML.
A: argeting the FLT3-ITD-F691L mutation has therapeutic implications for the treatment of AML. Tyrosine kinase inhibitors (TKIs) that specifically target the FLT3 protein, such as midostaurin (Rydapt), can be used to treat patients with this mutation. These drugs aim to inhibit the aberrant signaling and slow or stop the progression of leukemia cells with FLT3-ITD-F691L.
A: The FLT3-ITD-F691L mutation is detected through various molecular diagnostic techniques, including reverse transcription polymerase chain reaction (RT-PCR), fluorescence in situ hybridization (FISH), and next-generation sequencing (NGS). These methods can identify the specific ITD and F691L mutations, allowing for accurate diagnosis and prognosis of patients with AML and this mutation.
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The Human FLT3-ITD-F691L Stable Cell Line-Ba/F3 delivers highly reproducible outcomes, ensuring that experimental results are consistent and trustworthy. This trait is crucial for validating research findings and contributes to the robustness of scientific studies
The Human FLT3-ITD-F691L Stable Cell Line-Ba/F3 offers good value for money, making it a cost-effective choice for budget-conscious researchers. Its affordability increases its accessibility and is likely to broaden its application in research labs worldwide.
The Human FLT3-ITD-F691L Stable Cell Line-Ba/F3 has garnered positive reviews from researchers due to its high quality and performance. Its utility in studying FLT3-ITD-related leukemia has been well received, establishing its reputation as a reliable tool in the scientific community.
Customers of the Human FLT3-ITD-F691L Stable Cell Line-Ba/F3 can expect a high level of post-purchase support. The provider's responsive and efficient customer service ensures that any questions or issues are resolved quickly, leading to a satisfying user experience.
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