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dc.contributor.authorRodriguez-Losada, Noela
dc.contributor.authorWendelbo, Rune
dc.contributor.authorGarcia-Fernandez, Maria
dc.contributor.authorPavia, Jose
dc.contributor.authorMartinez-Montañez, Elisa
dc.contributor.authorLara-Muñoz, José Pablo 
dc.contributor.authorArenas, Ernest
dc.contributor.authorAguirre-Gomez, Jose Angel 
dc.date.accessioned2014-11-26T12:13:14Z
dc.date.available2014-11-26T12:13:14Z
dc.date.created2014-11-25
dc.date.issued2014-11-26
dc.identifier.urihttp://hdl.handle.net/10630/8489
dc.description.abstractThe emerging carbon nanomaterial Graphene (G), in the form of scaffold structure, has an efficient bioconjugation with common biomolecules and activates cell differentiation of neuronal stem cells, providing a promising approach for neural regeneration. We propose the use of G as a scaffold to re-address the dopaminergic (DA) neurons and the residual axons from dead or apoptotic DA neurons in Parkinson´s disease (PD). G could act as a physical support to promote the axonal sprout as a “deceleration” support for the DA cells derived from neural stem cells or DA direct cell conversion, allowing the propagation of nerve impulses. We cultured a clonal substantia nigra (SN) DA neuronal progenitor cell line (SN4741) in presence of G as scaffold. This cell line derived from mouse embryos was cultured in Dulbecco’s modified Eagle’s medium/10% FCS to about 80% confluence. Cells were incubated in three chemically different G derivatives and two different presentation matrixes as powder and films: 1) G oxide (GO); 2) partially reduced GO (PRGO) which is hydrophobic; and 3) fully reduced GO (FRGO). Cell viability was determined using the MTT assay after adding the following G concentrations: 1mg/ml; 0.1mg/ml; 0.05mg/ml; 0.02mg/ml and 0.01mg/ml, in each type of GO. To study cellular morphology and assessment of cell engraftment into GO films (GO film, PRGO film, FRGO film), we analyzed the immunostaining of the anti-rabbit neuron-specific DNA-binding protein (NeuN) antibody, the anti-rat Beta-3-tubulin antibody in combination with the mitochondrial marker mouse anti-ATP synthase antibody, and the anti-rabbit DCX as immature neuronal marker. Hoechst label was used as nuclei marker. Reactive oxidative species (ROS) were measured by flow cytometry to study the influence of G on the cell redox-state. With this purpose, cells were loaded with dihydroethidium. The mitochondrial membrane potential after JC-1 incubation was studied by flow cytometry. Our results show an increase of survival and metabolism (30-40%) at low concentrations of PRGO and FRGO (0.05-0.01 mg/ml) respect to the higher concentration (1 mg/ml), while no changes were seen in the GO group. LDH concentration was measured in the supernatant using a COBAS analyzer showing a neuroprotective action at low concentrations. Furthermore, either PRGO film or FRGO film show an increase in the effective anchorage capacity to nest into the G matrix and in the maturation of the SN 4741 cells. We conclude that the use of G scaffolds in the research of neurological diseases like PD could offer a powerful platform for neural stem cells, direct cell conversion techniques and neural tissue engineering.es_ES
dc.description.sponsorshipUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Norwegian Research Council (grant nº 215086)es_ES
dc.language.isoenges_ES
dc.rightsinfo:eu-repo/semantics/openAccesses_ES
dc.subjectParkinson, Enfermedad dees_ES
dc.subject.otherGraphenees_ES
dc.subject.otherStem cellses_ES
dc.subject.otherDopaminergices_ES
dc.subject.otherDifferentiationes_ES
dc.subject.otherParkinson's diseasees_ES
dc.titleGraphene derivative scaffolds facilitate in vitro cell survival and maturation of dopaminergic SN4741 cellses_ES
dc.typeinfo:eu-repo/semantics/conferenceObjectes_ES
dc.centroFacultad de Medicinaes_ES
dc.relation.eventtitle44th Annual Meeting of the Society for Neuroscience USA, Washintong DC, USAes_ES
dc.relation.eventplaceWashington DC, Estados Unidos de Americaes_ES
dc.relation.eventdate15-19 Noviembre 2014es_ES


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