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Additional file 1. Supplementary Figures of Manuscript. Supplementary tables and figures required for the better comprehension of the manuscript.

Being a multi-layered organ, the complexity of human skin is difficult to mimic in vitro. Despite the major advances in currently available 3D skin models, they remain very simplistic in relation to native tissue complexity. Moreover, the integration of melanoma cells into 3D skin models rendering representative tumor microenvironments is yet to be fully achieved. Cell sheet engineering has shown impressive results in recreating the native tissue structure and interactions of many different tissues by keeping intact the native cell-cell and cell-extracellular matrix interactions (Yanf et al., 2007; Haraguchi et al., 2012). Considering these major features, we built a 3D full skin model by stacking heterotypic cell sheets of human dermal fibroblasts (hDFbs) and keratinocytes (hKCs) or of hDFbs and human dermal microvascular endothelial cells (hDMECs), respectively as units of the epidermis- or dermis-like analogues. Melanocytes were further incorporated in the epidermis-like cell sheets before exposed to air-liquid interface (ALI), obtaining a pigmented stratified epidermis. In order to understand how melanoma cells responded to the self-organized full skin model, different cell lines (WM-115, SK-Mel-28, LM-MEL-33 and VMM15) were used. Histologically analysis confirmed the organization of those cells within the epidermis and their migration into the dermis through the formed basement membrane. Interactions between the melanoma cells and the stromal cells in the dermis are being assessed focusing on the known mechanisms involved in tumour progression in order to validate our approach., Acknowledgments: Authors acknowledge the financial support from ERC Consolidator Grant â ECM_INK (ERC-2016-COG-726061)

Additional file 1: Fig. S1. Phenotype of the CD49fbri/CD71dim subpopulation. A) Representative image of fluorescent activated cell sorting (FACS) dotplot of the whole population of epidermal keratinocytes and the gatting of the subpopulations of cells, based on the expression of the CD49f and CD71 markers, accompanied by the respective cellular percentages. CD49fbri/CD71dim cells represent epidermal stem-like cells, CD49fbri/CD71bri subpopulation are transit-amplifying (TA) cells, while differentiated (Diff.) cells are characterized by a CD49fdim phenotype. Epidermal stem-like cells derived colonies growing in the inactivated feeders (B) 4 and (C) 6 days after seeding. Data shown are mean ± SEM. Scale bars are 50 μm. Fig. S2. Human cell detection within skin explants from the hair reconstitution assay. Staining with a human-specific probe demonstrates that, 6 weeks after cells injection, there were no human cells remaining in the wound area in the controls (A,B) and in the condition where interfollicular epidermal stem-like keratinocytes (EpSlKCs) and DP cells were co-grafted (C). Likewise, no staining was observed in the mice tissue (D), whereas nuclear orange-pink staining was observed in human positive control specimens (E,F), demonstrating the specificity of the staining. Scale bars are 200 μm, for (A-D) and 50 μm for (E,F). Table S1. List of antibodies used for flow cytometry studies. Table S2. List of antibodies used for immunofluorescence studies.

Poster.-- Aquaculture Europe 2021, Funchal, Madeira, 4-7 October, Phosphorus is an essential nutrient for skeletal physiology, ionic homeostasis, and energy metabolism in fish. Unlike other minerals, the phosphorus content in water is insufficient to meet fish requirements and therefore the diet is the main external source of phosphorus. In aquaculture, commercial diets commonly have higher phosphorus concentration than the basal requirements in most fish species to ensure growth and prevent skeletal disorders. This practice poses a serious environmental problem as excess phosphorus in water from non-eaten food and urinary/fecal excretions produces eutrophication of aquatic ecosystems. Our previous studies with acellular-boned fish (Suarez-Bregua et al., 2021) revealed that a lower phosphorous concentration diet is possible without affecting the fish health and growth. In this study, we wondered if this is also applicable to cellular-boned fish species. Thus, the objective of this study was to analyze the effect of dietary phosphorus concentrations (low, standard, and high phosphorus diets) on bone mineralization of zebrafish (Danio rerio), a popular model organism and a cellular-boned teleost species, This work was funded by the Spanish Economy and Competitiveness Ministry projects AGL014-52473R and AGL2017-89648P to J. Rotllant. L. Guerrero-Peña was supported by pre-doctoral fellowship of the Spanish Personnel Research Training Program funded by Spanish Economy and Competitiveness Ministry (PRE2018-085475). L. Méndez-Martínez, was supported by pre-doctoral fellowship of the Xunta de Galicia (IN606A-2020/006)

Devido ao aumento de lesões associadas ao envelhecimento da população, a regeneração do tecido ósseo tem sido alvo de estudo. Apesar da vasta investigação neste sentido, os auto-enxertos continuam a prevalecer como tratamento de primeira linha, apesar das suas limitações. A extração de compostos de recursos marinhos para uso em abordagens de engenharia de tecidos emerge como uma alternativa promissora para regeneração de lesões ósseas. Neste capítulo apresenta-se um biomaterial promissor para aplicação em engenharia de tecidos duros tendo como base uma estratégia de valorização de sub-produtos marinhos, nomeadamente pele e dentes de tubarão., Os autores agradecem o apoio financeiro recebido da União Europeia através do Programa INTERREG—POCTEP, no âmbito dos Projetos 0687_NOVOMAR_1_P e 0245_IBEROS_1_E, através do Programa de cooperação transnacional Espaço Atlântico, no âmbito do Projeto MARMED (2011-1/164) e através do 7º Programa-Quadro de Investigação e Desenvolvimento Tecnológico (FP7), através do Projeto POLARIS (REGPOT-CT2012-316331). Os autores gostariam de agradecer também ao Centro Tecnológico del Mar (CETMAR, Vigo, Espanha) e COPEMAR SA (Espanha) pelo fornecimento dos subprodutos de tubarão. G.S.D agradece ao Programa Norte2020 (Portugal2020) pela bolsa de doutoramento (NORTE-08-5369-F SE-000044) e R.P. agradece à Fundação para a Ciência e a Tecnologia pelo contrato IF/00347/2015.