Immortalized Mouse EGFR Mutant M3^-/- Epidermal Keratinocytes

The Immortalized Immortalized Mouse EGFR Mutant M3^-/- Epidermal Keratinocytes were derived from primary keratinocytes of newborn EGFR null mice. The primary keratinocytes were immortalized in subsequent cultures after passaging for an extended period. The immortalized cells display similar phenotype to the primary keratinocytes including the cobblestone-shape morphology. The Immortalized Mouse EGFR Mutant M3^-/- Epidermal Keratinocytes are non-tumorigenic. They also differentiate with increased calcium, maintain a relatively stable karyotype no more aberrant than cultured primary keratinocytes, and produce epidermis upon orthotopic grafting onto nude mouse hosts. The M3 muscarinic acetylcholine receptor is knocked out in these cells. Upon introduction of v-ras^Ha oncogene, the Immortalized Mouse EGFR Mutant M3^-/- Epidermal Keratinocytes produce squamous cell carcinoma in orthotopic skin grafts. These cells are useful in studies focusing on epidermal keratinocyte biology and pathology.

Immortalization Method: Immortalization in subsequent cultures

BioSafety Level: II

Species: Mouse

Source Organ: Skin

Growth Properties: Adherent

Morphology: Cobble-stone

Population Doubling: N/A

Seeding Density: 20,000 - 30,000 cells/cm².

Markers: Keratin 14, Keratin 6

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow XI medium available at abm, Cat. No. TM011. To make the complete growth medium, add the following components to the base medium: 8% chelexed fetal bovine serum (TM999)*, Penicillin/Streptomycin Solution (G255) to a final concentration of 1% and 5 ng/ml recombinant human FGF7 (Z100595).
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: Immunofluorescence analysis of Keratin markers

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C