Influence of Fucoxanthin on Proliferative Activity of Human Melanocyte Culture
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Cell Technologies in Biology and Medicine, No. 2, August, 2020
Influence of Fucoxanthin on Proliferative Activity of Human Melanocyte Culture E. V. Dzhussoeva1, A. A. Gorkun1,2, I. M. Zurina1,2, N. V. Kosheleva1,2,3, T. D. Kolokol’tsova1,2, and I. N. Saburina1,2 Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 2, pp. 139-142, June, 2020 Original article submitted October 16, 2019 We studied the effect of algae pigment fucoxanthin on proliferative activity of melanocyte culture from human skin. Fucoxanthin in high concentrations can be cytotoxic, which was confirmed by changes in melanocyte morphology and a decrease in their proliferative activity. Key Words: melanocytes; fucoxanthin; drug testing; cytotoxicity; proliferative activity Melanocytes play a leading role in protecting the skin from the damaging effects of UV light by providing the production of pigment in the complex and multistage melanogenesis process. The search for new drugs that can affect different stages and levels of melanin synthesis regulation for suppression of hyperpigmentation and abnormal melanogenesis is in progress. Cell cultures are a unique model for testing of these products [1-3,10]. Algae pigment fucoxanthin is considered as a promising drug capable of modulating melanogenesis and producing multiple protective effects in various pathologies [6,7]. Our aim was to study the effect of different concentrations of fucoxanthin on proliferative activity of human skin melanocyte culture.
MATERIALS AND METHODS Stock and working solutions of fucoxanthin. Fucoxanthin powder (Anhui) was dissolved in DMEM/F-12 nutrient medium (BioloT) to a concentration of 500 µM (stock solution) and sterilized by filtering through a 0.22-µ filter (Millipore). For the experiments, the stock solution of fucoxanthin was diluted with the growth medium to concentrations of 250, 50, and 5 µM. Research Institute of General Pathology and Pathophysiology; 2Russian Medical Academy of Continuous Professional Education, Ministry of Health of the Russian Federation; 3Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, Russia. Address for correspondence: [email protected]. T. D. Kolokol’tsova 1
Human skin melanocyte culture. Primary culture of human skin melanocytes (CELL Applications, Inc.) that was delivered to the laboratory in a cryopreserved state was quickly thawed at 37°C, transferred to 15-ml centrifuge tubes, Hanks solution (PanEco) was added to the cell suspension to a volume of 5-7 ml for dilution of the preserving solution, and the tubes were centrifuged (7 min, 1000 rpm, 100g). The supernatant was removed, the cells were resuspended in complete growth medium for melanocytes (CELL Applications, Inc.) and transferred to Petri dishes (seeding density 104 cells/cm2). After attaining 80-85% confluence, the cells were harvested with Versene and 0.25% trypsin (PanEco), the cell concentration was brought to 104/ cm2 with complete medium, and the cells were plated on a new Petri dishes. The medium was replaced every 2 days. Analy
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