A Novel and Effective Method for Human Primary Skin Melanocytes and Metastatic Melanoma Cell Isolation

SIMPLE SUMMARY: The present paper describes a simple, non-enzymatic and effective method of melanocyte or metastatic melanoma cell isolation from skin or lymph node explants, respectively. The method is based on selective harvesting of melanocytes or melanoma cells emigrating from the explants. Thus, isolated cells display specific phenotypical and functional features of melanocytes/melanoma cells such as tyrosinase and Melan-A expression and melanin production. Furthermore, melanocyte or melanoma cell cultures are not contaminated by keratinocytes and/or fibroblasts. The method appears to be a useful tool for studies on the biology of melanocytes and malignant melanoma. ABSTRACT: The development of an effective method of melanocyte isolation and culture is necessary for basic and clinical studies concerning skin diseases, including skin pigmentation disorders and melanoma. In this paper, we describe a novel, non-enzymatic and effective method of skin melanocyte and metastatic melanoma cell isolation and culture (along with the spontaneous spheroid creation) from skin or lymph node explants. The method is based on the selective harvesting of melanocytes and melanoma cells emigrating from the cultured explants. Thereby, isolated cells retain their natural phenotypical features, such as expression of tyrosinase and Melan-A as well as melanin production and are not contaminated by keratinocytes and fibroblasts. Such melanocyte and melanoma cell cultures may be very useful for medical and cosmetology studies, including studies of antitumor therapies.