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

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.

Canine melanocytes: Immunohistochemical expression of melanocytic markers in different somatic areas

“…Melan A is considered one of the best and most specific markers for melanocytes in different species, dogs included 7,25–27 . Results from our study confirm the specificity and the usefulness of Melan A for the recognition of intraepidermic melanocytes, both in immunohistochemical and immunofluorescence evaluation.…”

“…[21][22][23][24] Melan A is considered one of the best and most specific markers for melanocytes in different species, dogs included. 7,[25][26][27] Results from our study confirm the specificity and the usefulness of Melan A for the recognition of intraepidermic melanocytes, both in immunohistochemical and immunofluorescence evaluation. Also, Melan A proved useful for the recognition of dermal melanocytes, particularly in the dermis of the chow chow, where the histiocytic/macrophagic origin of the cells also was ruled out using IBA1.…”

Anandamide-Modulated Changes in Metabolism, Glycosylation Profile and Migration of Metastatic Melanoma Cells

“…With the tumor progression, the protein glycosylation on cells changes from short and lowly-branched surface glycans (such as on the WM35 cells of the primary RGP site) to long and highly branched oligosaccharides (present on the A375-P metastatic cells), thus the lectin Con A affinity for metastatic cells is higher than for the primary cells and the glycan viscoelastic index significantly increases. Because melanoma tumors are highly heterogenic, melanoma cells isolated from the same patient often differ in metastatic potential, for example, established melanoma subcultures presented a varied doubling time [75], as well as disparate cell elasticity (AFM analysis) and glycosylation profile (QCM-D analysis; [36]). However, all results were consistent: MM7 and MM9 populations were the quickest to divide (low value of doubling time), less resistant to deformation (low E values) and with high metastatic potential (low K D value and high gVI value).…”

Preparation and characterization of novel nanostructured lipid carriers (NLC) and solid lipid nanoparticles (SLN) containing coenzyme Q10 as potent antioxidants and antityrosinase agents

“…About 10 5 melanoma cells were cultured in each well of 12 well plates and treated with Q10-NLC/SLN at a concentration of 50 μM for 24 h. Then, they were floated by trypsinization and washed with PBS, then, NaOH at a concentration of 2 M was added and and were incubated at 100 °C for 30 min. After incubation at 60 °C for 2 h, the melanin content in treated cells was measured at 405 nm [ [38] , [39] , [40] , [41] ].…”