title: Long-term culture of pluripotent stem-cell-derived human neurons on diamond--A substrate for neurodegeneration research and therapy.
abstract: Brain Computer Interfaces (BCI) currently represent a field of intense research aimed both at understanding neural circuit physiology and at providing functional therapy for traumatic or degenerative neurological conditions. Due to its chemical inertness, biocompatibility and stability, diamond is currently being actively investigated as a potential substrate material for culturing cells and for use as the electrically active component of a neural sensor. Here we provide a protocol for the differentiation of mature, electrically active neurons on microcrystalline synthetic thin-film diamond substrates starting from undifferentiated pluripotent stem cells. Furthermore, we investigate the optimal characteristics of the diamond microstructure for long-term neuronal sustainability. We also analyze the effect of boron as a dopant for such a culture. We found that the diamond crystalline structure has a significant influence on the neuronal culture unlike the boron doping. Specifically, small diamond microcrystals promote higher neurite density formation. We find that boron incorporated into the diamond does not influence the neurite density and has no deleterious effect on cell survival.
authors.affiliation: Regenerative Medicine Laboratory, School of Clinical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom; School of Chemistry, University of Bristol, Bristol BS8 1TS, United Kingdom.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Nistor
authors.affiliation: School of Chemistry, University of Bristol, Bristol BS8 1TS, United Kingdom. Electronic address: paul.may@bristol.ac.uk.
authors.forename: Paul W
authors.initials: PW
authors.lastname: May
authors.affiliation: The Hatherly Labs, School of Medical Sciences, University of Exeter, Exeter EX4 4EA, United Kingdom.
authors.forename: Francesco
authors.initials: F
authors.lastname: Tamagnini
authors.affiliation: The Hatherly Labs, School of Medical Sciences, University of Exeter, Exeter EX4 4EA, United Kingdom; Department of Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD, United Kingdom.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Randall
authors.affiliation: Regenerative Medicine Laboratory, School of Clinical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom; Trinity College Institute of Neuroscience, Lloyd Institute, Trinity College, Dublin 2, Ireland. Electronic address: maeve.caldwell@bristol.ac.uk.
authors.forename: Maeve A
authors.initials: MA
authors.lastname: Caldwell
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods*; D001672:Biocompatible Materials / Q000138:chemical synthesis; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D008422:Materials Testing; D058612:Nanodiamonds / Q000737:chemistry* / Q000648:ultrastructure; D019636:Neurodegenerative Diseases / Q000473:pathology / Q000628:therapy; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D010316:Particle Size; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology*; D013499:Surface Properties
chemical_list: D001672:Biocompatible Materials; D058612:Nanodiamonds
keywords: Boron-doped diamond; Diamond substrate; Long term culture; Neurodegeneration research; Neuronal differentiation; Neuronal survival; Pluripotent stem cells