Cat.No. : CSC-RO0238 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-RO0238 |
| Description | This cell line stably expresses EML4-ALK fusion gene with G1202R point mutation in the ALK part. |
| Target Gene | EML4 |
| 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 | EML4 echinoderm microtubule associated protein like 4 [ Homo sapiens ] |
| Gene Symbol | EML4 |
| Synonyms | EML4; echinoderm microtubule associated protein like 4; C2orf2; echinoderm microtubule-associated protein-like 4; ELP120; ROPP120; ropp 120; restrictedly overexpressed proliferation-associated protein; EMAP-4; EMAPL4; FLJ10942; FLJ32318; DKFZp686P18118; |
| Gene ID | 27436 |
| UniProt ID | Q9HC35 |
| mRNA Refseq | BC008685 |
| Chromosome Location | 2p21 |
| Function | molecular_function; |
| MIM | 607442 |
| 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 |
In first-line settings, alectinib has demonstrated superior efficacy in ALK-rearranged non-small-cell lung cancers. A Human EML4-ALK-G1202R Stable Cell Line was established to investigate acquired resistance mechanisms in ALK-rearranged non-small-cell lung cancers. This model enables the exploration of resistance mutations, such as ALK-I1171N and G1202R, frequently observed following treatment with alectinib, ceritinib, or lorlatinib. Through drug screening and computational simulations, potential therapeutic strategies for overcoming resistance were identified, underscoring the significance of this stable cell line in elucidating drug resistance mechanisms and developing effective treatment approaches for relapsed patients.
Figure 1. Resistance mechanisms in alectinib-refractory patients were investigated using Human EML4-ALK-G1202R Stable Cell Line. Cell viability was assessed upon treatment with various ALK inhibitors, providing insights into potential therapeutic strategies. (Okada K, et al., 2019)
A: Ba/F3 cells were likely chosen for their cytokine-dependent growth and suitability for studying oncogenic kinase activity and drug resistance mechanisms associated with EGFR mutations, including L858R, T790M, and C797S.
A: Stability was likely confirmed through methods such as immunoblotting, functional assays measuring downstream signaling, or cell viability assays in the absence of growth factors, with continuous selection pressure applied.
A: Characterization may involve analysis of EGFR phosphorylation, downstream signaling pathways, and functional implications in cell proliferation, survival, and response to EGFR inhibitors such as osimertinib or third-generation inhibitors targeting the T790M/C797S mutations.
A: Quality control likely included confirmation of EGFR-L858R/T790M/C797S expression levels, validation of its kinase activity and drug sensitivity, assessment of off-target effects, and validation of phenotypic changes associated with EGFR modulation.
A: Comparative analysis with patient-derived samples or in vivo models helps validate the relevance of EGFR-L858R/T790M/C797S expression in NSCLC progression, metastasis, and response to EGFR-targeted therapies, guiding the development of personalized treatment strategies for patients with acquired resistance mutations.
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Unrivaled reliability! The EML4-ALK-G1202R Stable Cell Line in Ba/F3 cells ensures stable expression of the ALK fusion protein, providing consistent results in cancer drug resistance studies.
Enabling advanced exploration! With stable EML4-ALK-G1202R expression, I can investigate mechanisms of resistance to ALK inhibitors with confidence, advancing precision medicine research in ALK-positive tumors.
Outstanding performance! This cell line exceeds expectations, serving as a robust platform for studying EML4-ALK-G1202R-targeted therapies and personalized treatment approaches in lung cancer.
Optimizing research workflows! Its stable expression simplifies experimental procedures, facilitating efficient data collection and analysis, and accelerating discoveries in drug resistance mechanisms.
An invaluable tool! The EML4-ALK-G1202R Stable Cell Line has significantly enhanced my research capabilities, offering valuable insights into ALK-driven oncogenesis and potential therapeutic strategies for overcoming drug resistance in ALK-positive cancers.
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