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

Aneta Ścieżyńska,Agata Góźdź,Marta Soszyńska,Krzysztof Łuszczyński,Tomasz Kobiela,Tomasz M Grzywa,Paweł K Włodarski,Ryszard Galus,Patrycja D Kowalska,Jacek Malejczyk,Anna Sobiepanek,Natalia Krześniak

doi:10.3390/cancers13246244

Abstract

Simple SummaryThe 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. 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.

Highlights

Considering the above, we focused on developing an alternative non-enzymatic method of skin melanocyte and metastatic melanoma cell isolation
Microscopic observation of skin explants revealed the emigration of cells usually starting from the second day of culture
The migrating cells formed pavement-like rings surrounding the skin explants (Figure 2A)

Summary

Introduction

Production of melanin, a natural pigment responsible for skin coloration. Melanocyte dysfunction can result in the loss of pigmentation, and appearance of hypoor hyperpigmented spots and may lead to the development of malignant melanoma [3]. Great progress has recently been made in the treatment of melanoma (e.g., owing to the ipilimumab and nivolumab therapy), there is still a high proportion of poorly responding patients [5]. This is due to a high malignancy and heterogenicity of melanoma cells that make the available therapies not fully effective [6,7]. Not surprisingly, melanoma is one of the most challenging cancers nowadays, in particular the cases of the refractory disease [8]

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