Cat.No. : CSC-RO0379 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-RO0379 |
| Description | This cell line is engineered to stably overexpress exogenous human FLT3 with ITD duplicate and D835Y mutations. |
| Target Gene | FLT3-ITD |
| 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 |
Autophagy plays a key role in drug resistance in acute myeloid leukemia (AML), including the FLT3-ITD mutant subtype. Here, researchers examined the expression of autophagy markers in FLT3-ITD-positive leukemia cells before and after acquired FLT3 inhibitor resistance. Researchers tested the stimulatory effects of the acquired D835Y mutation and the bone marrow microenvironment (BME) on autophagy and explored the mechanisms by which autophagy mediates FLT3 inhibitor resistance. These results showed that sorafenib-resistant cells significantly overexpressed autophagy markers compared with sorafenib-sensitive cells or cells before sorafenib treatment. Both the acquired D835Y mutation and BME activated cytoprotective autophagy to mediate FLT3 inhibitor resistance. Autophagy activation reduced the inhibitory effect of FLT3 inhibitors on FLT3 downstream signaling, thereby weakening their anti-leukemic effects. Inhibiting autophagy with CQ can significantly enhance the inhibitory effect of FLT3 inhibitors on FLT3 downstream signaling, ultimately overcoming resistance to FLT3 inhibitors.
To examine the activation of autophagy in sorafenib-resistant AML cells, researchers established sorafenib-resistant cell lines, such as Baf3 cells with FLT3-D835Y or FLT3-ITD + D835Y mutations, and examined the expression of autophagy markers, including LC3B, Beclin-1, ATG5, and p62. Consistent with previous reports, Ba/F3-FLT3-ITD + D835Y mutant cells were resistant to sorafenib (Figure 1A). Compared with sorafenib-sensitive Ba/F3-ITD mutant cells ( Figure 1A ), both Ba/F3-FLT3-D835Y and Ba/F3-FLT3-ITD + D835Y mutant cells showed higher LC3B-II, Beclin-1, and ATG5 expression and p62 degradation ( Figure 1B ), suggesting that sorafenib-resistant FLT3-ITD-positive cell lines can also overexpress autophagy and that cytoprotective autophagy may be activated by acquired resistance mutations.
Figure 1. Sorafenib-resistant cell lines showed overpresentation of autophagy markers. (Xu D, et al. 2022)
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
The Human FLT3-ITD-D835Y Stable Cell Line - BaF3 has significantly accelerated the pace of our research. The stability and consistency of this cell line allows us to generate reproducible data across multiple experiments, aiding drug development research with amazing efficiency.
The BaF3 cells expressing the FLT3-ITD-D835Y mutation provide an excellent model for mimicking the disease environment seen in acute myeloid leukemia (AML) patients. We've been able to test numerous compounds with reliable results, making this a crucial tool in our therapeutic screening process.
As a researcher focused on studying cancer signaling pathways, the Human FLT3-ITD-D835Y Stable Cell Line - BaF3 has proven to be indispensable. Its robustness and high transfection efficiency have allowed us to delve deeper into the molecular mechanisms that control FLT3 mutations.
Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.
Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Copyright © Creative Biogene. All rights reserved.
