Cultivation of Melanoma Cells in vitro on a 3D Scaffold Prepared on the Basis of Gelatin
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ivation of Melanoma Cells in vitro on a 3D Scaffold Prepared on the Basis of Gelatin A. A. Yatsenkoa, b, *, V. A. Kushnarevb, c, E. M. Ustinova, D. V. Leonova, V. M. Kislitskiya, S. S. Tseluykoa, and A. S. Artemievac a
Amur State Medical Academy of the Ministry of Health of the Russian Federation, Blagoveshchensk, 675006 Russia b LLC IQ Biofabric, Skolkovo, Moscow, 121205 Russia c National Medical Research Center of Oncology Named after N.N. Petrov of the Ministry of Health of the Russian Federation, St. Petersburg, Pesochny, 191124 Russia *e-mail: [email protected] Received October 1, 2019; revised February 10, 2020; accepted February 19, 2020
Abstract—Organotypic cultivation of melanoma cells in a three-dimensional matrix that mimics the dermis is a promising area of investigation for modeling a tumor and studying its biology. The aim of the study was to study the morphological and immunohistochemical characteristics of melanoma cells obtained as a result of an excision skin biopsy during prolonged 3D cultivation on a gelatin matrix. After 35 days of cultivation, light-optical morphological and immunohistochemical analysis of scaffolds was performed using antibodies to the melanocytic antigens S100, SOX10, as well as to the membrane antigen CD44. After 35 days of culturing the cells of the primary melanoma culture on porous gelatin scaffolds obtained by the leaching method, the expression of melanocytic antigens and membrane antigen CD44 was preserved. Melanoma cells adhered to the surface and destroyed the scaffold matrix. Thus, the component and quantitative composition of a three-dimensional gelatin scaffold has been developed, which ensures the adhesion and proliferation of cells of the primary melanoma culture while maintaining the expression of melanocytic antigens and protein CD44. Keywords: melanoma model in vitro, 3D cultivation, scaffold DOI: 10.1134/S1990519X20060097
Melanoma is the most aggressive and deadly form of skin tumor. The overall 5-year survival rate for patients with distant metastases is 16%, and the median overall survival period is 4–6 months (Faruk, 2012; Cheng et al., 2018). Until recently, treatment options have included attempts at chemotherapy and immunotherapy; however, with the discovery of the BRAF gene characteristic of 40–60% melanomas in activating mutations in codon 600, active use of its inhibitors began (Hayward et al., 2017; Luke et al., 2017). Rapid progress in the creation and development of this class of drugs is associated with the availability of suitable research models. The interaction of tumor cells with the microenvironment plays an important role in their functioning and regulation, and, therefore, is a critical factor in the response to therapy. The tumor matrix is one of the defining components of the microenvironment. Melanoma cells have the ability to invade the surrounding stroma, proliferate, and migrate (Shain and Bastian, 2016; Müller and Kulms, 2018). The effectiveness of the applied melanoma model depends on whether it preserves the morphology and a
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