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Abnormalities in the division of melanocytes are quite common, with potentially oncogenic growth usually followed by cell senescence producing benign naevi (moles), or occasionally, melanoma. Therefore, melanocytes are a useful model for studying both cellular senescence and melanoma, as well as many other aspects of biology such as pigmentation, organelle biogenesis and transport, and the diseases affecting these mechanisms. Melanocytes for use in basic research can be obtained from a range of sources, including surplus postoperative skin or from congenic murine skin. Here we describe methods to isolate and culture melanocytes from both human and murine skin (including the preparation of mitotically inactive keratinocytes for use as feeder cells). We also describe a high\u2010throughput transfection protocol for human melanocytes and melanoma cells. \u00a9 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Primary explantation of human melanocytic cells<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Preparation of keratinocyte feeder cells for use in the primary culture of mouse melanocytes<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Primary culture of melanocytes from mouse skin<\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: Transfection of human melanocytes and melanoma cells<\/jats:p>","DOI":"10.1002\/cpz1.774","type":"journal-article","created":{"date-parts":[[2023,5,8]],"date-time":"2023-05-08T12:10:29Z","timestamp":1683547829000},"source":"Crossref","is-referenced-by-count":13,"title":["Isolation, Culture, and Transfection of Melanocytes"],"prefix":"10.1002","volume":"3","author":[{"given":"Philip S.","family":"Goff","sequence":"first","affiliation":[{"name":"Cell Biology Research Section, Molecular and Clinical Sciences Research 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