Epidermal melanocytes are melanin-producing cells that function as a protective mechanism from harmful UV rays. Melanoma has been associated to defects, dysfunction, or lack of melanocytes. Primary C57BL/6 mouse melanocytes have been derived and successfully immortalized with SV40 large T antigen. The Immortalized Mouse Late Stage Melanocytes (iMC65) are hygromycin-resistant cells which expresses melanogenic markers (c-kit, Pax3, Sox10, and MITF-m), and are non-tumorigenic. The mature melanocytes express HMB45 and high tyrosinase activity. These cells are a valuable tool in melanocyte biology and molecular pathogenesis of pigment-related cell disorder studies.
Immortalization Method: Serial passaging and transduction with retrovirus SSR#69 expressing SV40 large T antigen flanked with Cre/loxP sites
BioSafety Level: II
Species: Mouse
Source Organ: Skin
Growth Properties: Adherent
Morphology: Polygonal
Population Doubling: N/A
Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers: Pax3, Sox10, MITF-m, c-kit
Donor Age: N/A
Donor Gender: N/A
Donor Ethnicity: N/A
Propagation: The base medium for this cell line is Prigrow III medium available at abm (TM003). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. 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: 1) Fontana-Masson staining for melanocytic cell markers; 2) Tyrosine enzymatic assays determined presence of tyrosine; 3) Western blot analysis for SV40 large T antigen presence
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C