EML4‑ALK mutant murine lung cancer cell line

Tool Details

*FOR RESEARCH USE ONLY (for other uses, please contact the licensing team)

• Name: EML4‑ALK mutant murine lung cancer cell line
• Alternate name: EML4‑ALK mutant cell line; Y143 cell line
• Cancer type: Lung cancer
• Research fields: Cancer biology; Immunotherapy
• Organism: Mouse
• Gender: Female
• Tissue: Lung
• Disease: Cancer
• Description: A novel syngeneic EML4-ALK mutant murine lung cancer cell line (Y143) with orthotopic growth potential, that can serve as a useful preclinical model for evaluating tumour-host interactions, the impact of specific oncogenic alterations on the tumour microenvironment, and immunotherapeutic approaches. This cell line was derived from the primary lung tumour of a female C57BL/6 mouse harbouring the EML4-ELK gene fusion and is capable of forming orthotopic tumours in immunocompetent C57BL/6 mice. Y143 cells demonstrate the EML4-ALK genetic rearrangement as shown by PCR. In Y143 cells treatment with the EML4-ALK inhibitor crizotinib inhibits phosphorylation of AKT and ERK is inhibited in a dose dependent manner and treatment with the ALK kinase inhibitor, TAE684, inhibits growth of Y143 cells.
• Application: 2D cell culture; Immunohistochemistry (IHC); Cell line-derived xenograft (CDX) model development
• Production details: An 8 week old C57BL/6 female mouse was treated with 30 µl of 10^6 PFU/ml Ad-EA by tracheal instillation. The Ad-EA vector has an Ad5 backbone and harbors Cas9 and guide RNAs that lead to the EML4-ALK gene fusion. When this animal was euthanized 14 week later a group of multifocal tumors 3–5 mm in size were combined and passaged into a recipient animal. For this line, tumors formed in both lung and flank; these tumors were combined and then passaged together into both lung and flank sites of recipient animals. Subsequently, the EML4-ALK mutant cell line (Y143) was established that was capable of forming orthotopic tumours in immunocompetent C57BL/6 animals.
• Biosafety level: 1

Applications

• Application: 2D cell culture; Immunohistochemistry (IHC); Cell line-derived xenograft (CDX) model development

Handling

• Format: Frozen
• Growth medium: Dulbecco's Modified Eagle Medium (DMEM) containing 4.5 g/l d-glucose, l-glutamine, and sodium pyruvate, supplemented with 10% (v/v) fetal bovine serum and 100 μg/ml of primocin (optional)
• Temperature: 37°C
• Atmosphere: 5% CO2
• Shipping conditions: Dry ice
• Storage conditions: Liquid Nitrogen
• Subculture routine: Split 90% confluent cultures 1:5 to 1:15 as required. Cells are cultured at 37°C, in a humidified incubator with 5% CO2.
• Cultured in antibiotics: Primocin (optional)

References

• Nolan et al. 2020. Cancer Cell Int. 28: 20:417. PMID: 32874131