KrasG12D.Map3k7−/−.GFP+ murine lung cancer cell line

Tool Details

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

• Name: KrasG12D.Map3k7−/−.GFP+ murine lung cancer cell line
• Alternate name: KrasG12D.Map3k7−/−.GFP+ cell line; E889 cell line
• Cancer type filter: Lung cancer
• Research fields: Cancer biology; Immunotherapy
• Organism: Mouse
• Gender: Male
• Tissue: Lung
• Disease: Cancer
• Description: A novel syngeneic KrasG12D.Map3k7-/-.GFP+ murine lung cancer cell line (E889) 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 tumor microenvironment, and immunotherapeutic approaches. This cell line was derived from the primary lung tumour of a KrasLSL-G12D/+.Map3k7fl/fl. ROSAmTmG male C57BL/6 mouse and is capable of forming orthotopic tumours in immunocompetent C57BL/6 mice. E889 cells demonstrate genetic recombination at the Kras and Map3k7 loci as shown by PCR, and express KRASG12D but not MAP3K7, as shown by western analysis. As this cell line was derived from an animal harboring the ROSAmTmG tracking allele, it also expresses GFP.
• Application: 2D cell culture; Immunohistochemistry (IHC); Cell line-derived xenograft (CDX) model development
• Production details: An 8 week old KrasLSL-G12D/+.Map3k7fl/fl. ROSAmTmG male mouse was injected with 2 µl of 10^9 PFU/ml Ad5-CMV-Cre into the left lung. When this animal was euthanized 26 week later, an 8 mm primary tumour was passaged into a male recipient. After two passages through the flanks of recipient animals, the KrasG12D.Map3k7-/-.GFP+ cell line (E889) was established that was capable of forming orthotopic tumors 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