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A photorealistic image of a science laboratory with a microscope and several cell culture plates on a desk.

I. Testes in vitro: Testes de viabilidade celular Foram realizados, em primeiro lugar, testes in vitro. Utilizámos as linhas celulares CCR (HCT116 e SW480), cultivadas num meio Eagle modificado por Dulbecco com 10 % de soro fetal bovino (FBS) e 1 % de penicilina. As células foram mantidas numa incubadora a 37 °C. As células foram semeadas em placas de 96 poços (5000 células) tratadas com uma solução à base de curcumina, uma solução à base de EGCG ou uma solução contendo ambos os compostos as diferentes concentrações. Em cada placa, foi também realizado um controlo positivo, tratando as células com DMSO, e um controlo negativo contendo apenas células em meio à cultura. As células foram mantidas durante 48 horas na incubadora . Foram realizados testes de citotoxicidade com resazurina em todas as células para medir a sua atividade metabólica. Apenas para as células tratadas com curcumina e combinação dos dois compostos : Para os testes seguintes, células da mesma linha celular foram tratadas durante 16 horas com soluções de curcumina e dos dois compostos. Em seguida, realizamos medições da quantidade de ROS, que se acumulam durante a ferroptose, tratando as células com DCFH-DA, uma molécula que penetra nas células e atravessa a membrana celular. A molécula é então ativada pelas enzimas celulares e oxidada pelos ROS, tornando-se fluorescente. A fluorescência é então medida com um microscópio de fluorescência. Quanto mais intensa for a fluorescência, maior será a quantidade de Mehr sehen

#science #laboratory #biotechnology

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