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Immortalized-Mouse-Melanocytes

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Immortalized-Mouse-Melanocytes

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	Print Version
BioSafety Level	II
Organism	Mouse
Source Organ	Skin
Growth Properties	Adherent
Morphology	Polygonal
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	c-kit, Pax3, Sox10, MITF-m
Applications	For Research Use Only
Immortalization Method	Serial passaging and transduction with retrovirus SSR#69 expressing SV40 large T antigen flanked with Cre/loxP sites
Description	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.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow III medium available from 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 (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C.
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
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.