Navegando por Autor "Rehen, Stevens K."
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Artigo Computational fluid dynamic analysis of physical forces playing a role in brain organoid cultures in two different multiplex platforms(2019-03-07) Goto-Silva, Livia; Ayad, Nadia M. E.; Herzog, Iasmin L.; Silva, Nilton P.; Lamien, Bernard; Orlande, Helcio R. B.; Souza, Annie da Costa; Ribeiro, Sidarta Tollendal Gomes; Martins, Michele; Domont, Gilberto B.; Junqueira, Magno; Tovar-Moll, Fernanda; Rehen, Stevens K.BACKGROUND: Organoid cultivation in suspension culture requires agitation at low shear stress to allow for nutrient diffusion, which preserves tissue structure. Multiplex systems for organoid cultivation have been proposed, but whether they meet similar shear stress parameters as the regularly used spinner flask and its correlation with the successful generation of brain organoids has not been determined. RESULTS: Here we used computational fluid dynamics (CFD) to simulate two multiplex culture conditions: steering plates on an orbital shaker and the use of a previously described bioreactor. The bioreactor had low speed and high shear stress regions that may affect cell aggregate growth, depending on volume, whereas the computed variables of the steering plates were closer to those of the spinning flask. CONCLUSION: Our protocol improves the initial steps of the standard brain organoid formation, and the produced organoids displayed regionalized brain structures, including retinal pigmented cells. Overall, we conclude that suspension culture on orbital steering plates is a cost-effective practical alternative to previously described platforms for the cultivation of brain organoids for research and multiplex testing.Artigo Short term changes in the proteome of human cerebral organoids induced by 5-MeO-DMT(2017-10-09) Dakic, Vanja; Nascimento, Juliana Minardi; Sartore, Rafaela Costa; Maciel, Renata de Moraes; Araújo, Dráulio Barros de; Ribeiro, Sidarta Tollendal Gomes; Martins-de-Souza, Daniel; Rehen, Stevens K.Dimethyltryptamines are entheogenic serotonin-like molecules present in traditional Amerindian medicine recently associated with cognitive gains, antidepressant effects, and changes in brain areas related to attention. Legal restrictions and the lack of adequate experimental models have limited the understanding of how such substances impact human brain metabolism. Here we used shotgun mass spectrometry to explore proteomic differences induced by 5-methoxy-N,N-dimethyltryptamine (5-MeO-DMT) on human cerebral organoids. Out of the 6,728 identified proteins, 934 were found differentially expressed in 5-MeO-DMT-treated cerebral organoids. In silico analysis reinforced previously reported anti-inflammatory actions of 5-MeO-DMT and revealed modulatory effects on proteins associated with long-term potentiation, the formation of dendritic spines, including those involved in cellular protrusion formation, microtubule dynamics, and cytoskeletal reorganization. Our data offer the first insight about molecular alterations caused by 5-MeO-DMT in human cerebral organoids.