{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T03:48:07Z","timestamp":1783828087900,"version":"3.55.0"},"reference-count":26,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2021,12,13]],"date-time":"2021-12-13T00:00:00Z","timestamp":1639353600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Center","doi-asserted-by":"publisher","award":["2016\/23\/N\/NZ5\/02588"],"award-info":[{"award-number":["2016\/23\/N\/NZ5\/02588"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004281","name":"National Science Center","doi-asserted-by":"publisher","award":["2017\/27\/N\/ST4\/01389"],"award-info":[{"award-number":["2017\/27\/N\/ST4\/01389"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Polish Ministry of National Defence","award":["523\/2017\/DA"],"award-info":[{"award-number":["523\/2017\/DA"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/cancers13246244","type":"journal-article","created":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T01:20:41Z","timestamp":1639444841000},"page":"6244","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["A Novel and Effective Method for Human Primary Skin Melanocytes and Metastatic Melanoma Cell Isolation"],"prefix":"10.3390","volume":"13","author":[{"given":"Aneta","family":"\u015acie\u017cy\u0144ska","sequence":"first","affiliation":[{"name":"Department of Histology and Embryology, Medical University of Warsaw, 02-004 Warsaw, Poland"},{"name":"Laboratory of Experimental Immunology, Military Institute of Hygiene and Epidemiology, 01-163 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3186-7582","authenticated-orcid":false,"given":"Anna","family":"Sobiepanek","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Interactions Studies, Faculty of Chemistry, Warsaw University of Technology, 00-661 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0635-7214","authenticated-orcid":false,"given":"Patrycja D.","family":"Kowalska","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Interactions Studies, Faculty of Chemistry, Warsaw University of Technology, 00-661 Warsaw, Poland"},{"name":"Polish Stem Cell Bank, 00-867 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Soszy\u0144ska","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Medical University of Warsaw, 02-004 Warsaw, Poland"},{"name":"Laboratory of Experimental Immunology, Military Institute of Hygiene and Epidemiology, 01-163 Warsaw, Poland"},{"name":"Doctoral School, Medical University of Warsaw, 02-091 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krzysztof","family":"\u0141uszczy\u0144ski","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Medical University of Warsaw, 02-004 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1809-248X","authenticated-orcid":false,"given":"Tomasz M.","family":"Grzywa","sequence":"additional","affiliation":[{"name":"Doctoral School, Medical University of Warsaw, 02-091 Warsaw, Poland"},{"name":"Centre for Preclinical Research, Department of Methodology, Medical University of Warsaw, 02-091 Warsaw, Poland"},{"name":"Department of Immunology, Medical University of Warsaw, 02-091 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Krze\u015bniak","sequence":"additional","affiliation":[{"name":"Medical Centre of Postgraduate Education, Department of Plastic and Reconstructive Surgery, Prof. W. Orlowski Memorial Hospital, 00-416 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Agata","family":"G\u00f3\u017ad\u017a","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Medical University of Warsaw, 02-004 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0683-1791","authenticated-orcid":false,"given":"Pawe\u0142 K.","family":"W\u0142odarski","sequence":"additional","affiliation":[{"name":"Centre for Preclinical Research, Department of Methodology, Medical University of Warsaw, 02-091 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryszard","family":"Galus","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Medical University of Warsaw, 02-004 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7155-659X","authenticated-orcid":false,"given":"Tomasz","family":"Kobiela","sequence":"additional","affiliation":[{"name":"Laboratory of Biomolecular Interactions Studies, Faculty of Chemistry, Warsaw University of Technology, 00-661 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5371-8476","authenticated-orcid":false,"given":"Jacek","family":"Malejczyk","sequence":"additional","affiliation":[{"name":"Department of Histology and Embryology, Medical University of Warsaw, 02-004 Warsaw, Poland"},{"name":"Laboratory of Experimental Immunology, Military Institute of Hygiene and Epidemiology, 01-163 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,13]]},"reference":[{"key":"ref_1","first-page":"013","article-title":"Cultivation of normal human epidermal melanocytes in the absence of phorbol esters","volume":"107","author":"Hsu","year":"2005","journal-title":"Methods Mol. Med."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Litwack, G. (2018). Chapter 13-Metabolism of amino Acids. Human Biochemistry, Academic Press.","DOI":"10.1016\/B978-0-12-383864-3.00013-2"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"540","DOI":"10.3389\/fbioe.2020.00540","article-title":"Human melanocyte-derived spheroids: A precise test system for drug screening and a multicellular unit for tissue engineering","volume":"8","author":"Zurina","year":"2020","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1056\/NEJMoa1910836","article-title":"Five-Year survival with combined nivolumab and ipilimumab in advanced melanoma","volume":"381","author":"Larkin","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"956","DOI":"10.1016\/j.tranon.2017.09.007","article-title":"Intratumor and intertumor heterogeneity in melanoma","volume":"10","author":"Grzywa","year":"2017","journal-title":"Transl. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1007\/s00249-021-01514-8","article-title":"Biophysical characterization of melanoma cell phenotype markers during metastatic progression","volume":"50","author":"Sobiepanek","year":"2021","journal-title":"Eur. Biophys. J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1007\/s11912-017-0575-8","article-title":"The role of surgery for melanoma in an era of effective systemic therapy","volume":"19","author":"Raigani","year":"2017","journal-title":"Curr. Oncol. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1016\/j.jfma.2018.02.010","article-title":"Characteristics of melanocyte spheroids formed through different biomaterial-induced processes","volume":"118","author":"Hsiao","year":"2019","journal-title":"J. Formos. Med. Assoc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1.8.1","DOI":"10.1002\/0471143030.cb0108s63","article-title":"Isolation, culture, and transfection of melanocytes","volume":"63","author":"Godwin","year":"2014","journal-title":"Curr. Protoc. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1007\/978-1-4939-9473-1_3","article-title":"Isolation and culture of epidermal melanocytes","volume":"Volume 1993","author":"Cario","year":"2019","journal-title":"Skin Tissue Engineering"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1111\/j.1600-0846.2011.00510.x","article-title":"Using human hair follicle-derived keratinocytes and melanocytes for constructing pigmented tissue-engineered skin","volume":"17","author":"Liu","year":"2011","journal-title":"Ski. Res. Technol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/s00403-007-0742-0","article-title":"Human adult epidermal melanocytes cultured without chemical mitogens express the EGF receptor and respond to EGF","volume":"299","author":"Szabad","year":"2007","journal-title":"Arch. Dermatol. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1111\/cpr.12028","article-title":"New procedure for epidermal cell isolation using kiwi fruit actinidin, and improved culture of melanocytes in the presence of leukaemia inhibitory factor and forskolin","volume":"46","author":"Yarani","year":"2013","journal-title":"Cell Prolif."},{"key":"ref_15","first-page":"107","article-title":"Isolation and culture of human primary keratinocytes\u2013A methods review","volume":"28","author":"Nogowska","year":"2018","journal-title":"Exp. Dermatol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1007\/s10616-018-0207-7","article-title":"Development and validation of a simple method for the extraction of human skin melanocytes","volume":"70","author":"Wang","year":"2018","journal-title":"Cytotechnology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1111\/j.1600-0749.1994.tb00016.x","article-title":"Organotypic culture of human skin to study melanocyte migration","volume":"7","author":"Wijngaard","year":"1994","journal-title":"Pigment. Cell Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"30","DOI":"10.5114\/pdia.2013.33376","article-title":"Skin melanocytes: Biology and development","volume":"30","author":"Cichorek","year":"2013","journal-title":"Postepy Dermatol. Alergol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"938","DOI":"10.1111\/j.1600-0625.2011.01356.x","article-title":"A new spectrophotometric method for simple quantification of melanosomal transfer from melanocytes to keratinocytes","volume":"20","author":"Kasraee","year":"2011","journal-title":"Exp. Dermatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1111\/j.1600-0625.2009.00900.x","article-title":"An improved method of human keratinocyte culture from skin explants: Cell expansion is linked to markers of activated progenitor cells","volume":"18","author":"Guo","year":"2009","journal-title":"Exp. Dermatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1111\/j.1600-0749.1996.tb00089.x","article-title":"Effective elimination of fibroblasts in cultures of melanocytes by lowering calcium concentration in TPA depleted medium following geneticin treatment","volume":"9","author":"Horikawa","year":"1996","journal-title":"Pigment. Cell Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"258","DOI":"10.4103\/ijdvl.IJDVL_275_17","article-title":"Melanocyte spheroids are formed by repetitive long-term trypsinization","volume":"85","author":"Zhang","year":"2019","journal-title":"Indian J. Dermatol. Venereol. Leprol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1038\/s41598-018-37055-y","article-title":"Quantitative analysis of melanin content in a three-dimensional melanoma cell culture","volume":"9","author":"Chung","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1016\/S0002-9440(10)63657-7","article-title":"MLANA\/MART1 and SILV\/PMEL17\/GP100 are transcriptionally regulated by MITF in melanocytes and melanoma","volume":"163","author":"Du","year":"2003","journal-title":"Am. J. Pathol."},{"key":"ref_25","first-page":"317","article-title":"Culture medium induced morphological changes of melanoma cells associated with change in sensitivity to lysis by lymphokine-activated killer cells","volume":"12","author":"Perng","year":"1997","journal-title":"Cancer Biother. Radiopharm."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1111\/j.1582-4934.2001.tb00175.x","article-title":"Basic studies on gene therapy of human malignant melanoma by use of the human interferon beta gene entrapped in cationic multilamellar liposomes. 1. Morphology and growth rate of six melanoma cell lines used in transfection experiments with the human interferon beta gene","volume":"5","author":"Benga","year":"2001","journal-title":"J. Cell. Mol. Med."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/13\/24\/6244\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:46:27Z","timestamp":1760168787000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/13\/24\/6244"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,13]]},"references-count":26,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["cancers13246244"],"URL":"https:\/\/doi.org\/10.3390\/cancers13246244","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,13]]}}}