Cat.No. : CSC-RO0414 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-RO0414 |
| Description | This cell line is engineered to stably overexpress exogenous human EPOR protein. |
| Target Gene | EPOR |
| 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 |
Cell surface receptors receive signals from outside the cell and play a crucial role in regulating various aspects of cell behavior by coordinating the cellular response. Researchers have explored the role of the human erythropoietin receptor (EPOR) in cell proliferation by using a 46-residue artificial transmembrane protein aptamer, ELI-3. The key role of EPOR in erythropoiesis and other cellular processes is achieved through the formation of EPOR homodimers or hetero-oligomers of EPOR with other receptors. However, the mechanisms of heteroreceptor assembly and signaling are unclear. The researchers found that ELI-3 was able to bind and activate EPOR, thereby triggering growth factor independence in EPOR-expressing mouse BaF3 cells. The activity of ELI-3 was dependent on the transmembrane region of EPOR and the JAK2-binding site, rather than cytoplasmic tyrosine, which is used for signaling through conventional EPOR. In contrast, ELI-3-induced proliferation and activation of JAK/STAT signaling required the transmembrane and cytoplasmic domains of the cytokine receptor β-common subunit (βcR). Furthermore, ELI-3 could not induce erythroid differentiation of human primary hematopoietic progenitors but inhibited serum withdrawal-induced nonhematopoietic cell death.
Figure 1. Researchers used YX4 expression libraries to screen for novel traptamers synergizing EPOR in BaF3/hEPOR cells. BaF3/hEPOR cells normally require IL-3 for proliferation, but EPO or proteins that activate hEPOR can substitute for EPO. Following the screen, these cells survived in media lacking growth factors, indicating that these traptamers make the cell's growth factor independent. (He L, et al., 2019)
A: This cell line, overexpressing erythropoietin receptor (EPOR), is ideal for bioassays to measure the potency and efficacy of EPO and its analogs by assessing STAT5 phosphorylation or cell proliferation in response to EPO stimulation.
A: Researchers can use this cell line to dissect the signaling pathways activated by EPO-EPOR binding, such as JAK2/STAT5, MAPK, and PI3K/Akt, by using specific pathway inhibitors and observing their effects on cell proliferation and differentiation.
A: Yes, this cell line is suitable for high-throughput screening assays to identify novel EPO-mimetic agents, as the overexpressed EPOR allows for a robust and measurable cellular response to EPO and its mimetics.
A: Regular validation by flow cytometry to detect surface expression of EPOR, along with Western blotting for total EPOR protein levels, is recommended to ensure stable expression over multiple passages.
A: This cell line can serve as a model to study the altered response to EPO in anemia of chronic disease by mimicking the conditions of impaired EPOR signaling and testing the efficacy of novel therapeutic agents designed to enhance EPO sensitivity or function.
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This cell line features stable overexpression of the human EPOR protein, which is essential for consistent experimental results in erythropoiesis and cancer research.
This cell line allows us to understand the role of EPOR in erythropoiesis and its potential involvement in cancer, providing a robust model for disease mechanism studies.
It is ideal for bioassays to measure the potency and efficacy of EPO and its analogs by assessing cellular responses to EPO stimulation.
The cell line can be used to model resistance development to EPO receptor inhibitors, facilitating the identification of resistance pathways in anemia treatment.
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