Immortalized Mouse Melanocytes

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 Melanocytes are hygromycin-resistant cells which expresses melanogenic markers (c-kit, Pax3, Sox10, and MITF-m), and are non-tumorigenic. The cells are a pool consisting of progenitor-like, intermediately-differentiated, and late-stage melanocytes. Individual clones are available: Immortalized Mouse Progenitor-Like Melanocytes (iMC23) (Cat. No. T0318), Immortalized Mouse Intermediately Differentiated Melanoblasts (iMC65) (Cat. No.T0320), and Immortalized Mouse Late Stage Melanocytes (iMC37)(Cat. No. T0319). By culturing the cells in the presence of dexamethasone, differentiation occurs and yields mature melanocytes expressing HMB45 and tyrosinase. 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: c-kit, Pax3, Sox10, MITF-m

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