TY - JOUR A1 - Gentile, Luca T1 - A complete workflow for the differentiation and the dissociation of hiPSC-derived cardiospheres N2 - Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) are an invaluable tool for both basic and translational cardiovascular research. The potential that these cells hold for therapy, disease modeling and drug discovery is hampered by several bottlenecks that currently limit both the yield and the efficiency of cardiac induction. Here, we present a complete workflow for the production of ready-to-use hiPSC-CMs in a dynamic suspension bioreactor. This includes the efficient and highly reproducible differentiation of hiPSCs into cardiospheres, which display enhanced physiological maturation compared to static 3D induction in hanging drops, and a novel papain-based dissociation method that offers higher yield and viability than the broadly used dissociation reagents TrypLE and Accutase. Molecular and functional analyses of the cardiomyocytes reseeded after dissociation confirmed both the identity and the functionality of the cells, which can be used in downstream applications, either as monolayers or spheroids. KW - hiPSCs KW - Cardiomyocytes KW - Cardiac induction KW - 3D biorector KW - Papain dissociation Y1 - 2018 U6 - https://doi.org/10.1016/j.scr.2018.08.015 VL - 2018 IS - 32/2018 SP - 65 EP - 72 PB - Elsevier ER - TY - JOUR A1 - Gentile, Luca T1 - Assessment of Iron Oxide Nanoparticle Ecotoxicity on Regeneration and Homeostasis in the Replacement Model System Schmidtea mediterranea JF - Altex N2 - Iron oxide nanoparticles (IONs) are used in a number of applications from food to cosmetics and from medical applications to magnetic storage. In spite of the 550 tons produced each year in Europe alone, no effective dose limit recommendations are established and the overall risks connected to IONs are still debated. The incorporation of IONs in daily life raises a concern about their effects on the environment, on living organisms, and on human health. In this study, we used freshwater planarians to assess the nanoecotoxicity of IONs. Planarians are free-living invertebrates known for their astonishing regenerative ability. Because of their sensitivity to toxicants, they are often used to determine the effects of toxic, genotoxic, and carcinogenic environmental compounds with an approach in line with the 3Rs (Reduce, Refine, Replace) principle. Planarians were exposed to IONs at concentrations up to 1 mg/ml and their effects were evaluated at the behavioral, morphofunctional, and molecular levels, with a special emphasis on the regeneration process. Our results indicate that IONs did not affect the stem cell population dynamics, nor did they induce substantial changes in either homeostatic or regenerating planarians. As positive controls, gold nanoparticles coated with the pro-apoptotic anti-cancer drug hexadecylmethylammonium bromide and highly concentrated polystyrene nanoparticles were used; these all elicited toxic effects. Therefore, we conclude that IONs at environmental concen- trations are safe for planarians, and that the planarian is a powerful model system that can replace vertebrate animal models in nanoecotoxicology research and for nanoecotoxicology studies. KW - 3R strategy KW - Pluripotency-based regeneration freshwater KW - Ecotoxicity KW - Reactive oxygen species Schmidtea mediterranea Y1 - 2019 U6 - https://doi.org/10.14573/altex.1902061 IS - 36(4)/2019 SP - 583 EP - 596 ER -