title: Generation of a Functional Human Neural Network by NDM29 Overexpression in Neuroblastoma Cancer Cells.
abstract: Recent advances in life sciences suggest that human and rodent cell responses to stimuli might differ significantly. In this context, the results achieved in neurotoxicology and biomedical research practices using neural networks obtained from mouse or rat primary culture of neurons would benefit of the parallel evaluation of the same parameters using fully differentiated neurons with a human genetic background, thus emphasizing the current need of neuronal cells with human origin. In this work, we developed a human functionally active neural network derived by human neuroblastoma cancer cells genetically engineered to overexpress NDM29, a non-coding RNA whose increased synthesis causes the differentiation toward a neuronal phenotype. These cells are here analyzed accurately showing functional and morphological traits of neurons such as the expression of neuron-specific proteins and the possibility to generate the expected neuronal current traces and action potentials. Their morphometrical analysis is carried out by quantitative phase microscopy showing soma and axon sizes compatible with those of functional neurons. The ability of these cells to connect autonomously forming physical junctions recapitulates that of hippocampal neurons, as resulting by connect-ability test. Lastly, these cells self-organize in neural networks able to produce spontaneous firing, in which spikes can be clustered in bursts. Altogether, these results show that the neural network obtained by NDM29-dependent differentiation of neuroblastoma cells is a suitable tool for biomedical research practices.
authors.affiliation: ETT Spa, via Sestri 37, 16154, Genoa, Italy.|National Research Council, Institute of Biophysics, via De Marini 6, 16149, Genoa, Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Alloisio
authors.affiliation: IRCCS-AOU San Martino-IST, Genova, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Garbati
authors.affiliation: National Research Council, Institute of Biophysics, via De Marini 6, 16149, Genoa, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Viti
authors.affiliation: Istituto Italiano di Tecnologia, Via Morego 30, I-16163, Genova, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Dante
authors.affiliation: IRCCS-AOU San Martino-IST, Genova, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Barbieri
authors.affiliation: IRCCS-AOU San Martino-IST, Genova, Italy.|Department of Experimental Medicine (DIMES), University of Genova, Largo Rosanna Benzi 10, 16132, Genova, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Arnaldi
authors.affiliation: Istituto Italiano di Tecnologia, Via Morego 30, I-16163, Genova, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Petrelli
authors.affiliation: IRCCS-AOU San Martino-IST, Genova, Italy.|Department of Experimental Medicine (DIMES), University of Genova, Largo Rosanna Benzi 10, 16132, Genova, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Gigoni
authors.affiliation: Department of Experimental Medicine (DIMES), University of Genova, Largo Rosanna Benzi 10, 16132, Genova, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Giannoni
authors.affiliation: IRCCS-AOU San Martino-IST, Genova, Italy.|Department of Experimental Medicine (DIMES), University of Genova, Largo Rosanna Benzi 10, 16132, Genova, Italy.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Quarto
authors.affiliation: National Research Council, Institute of Biophysics, via De Marini 6, 16149, Genoa, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Nobile
authors.affiliation: National Research Council, Institute of Biophysics, via De Marini 6, 16149, Genoa, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Vassalli
authors.affiliation: IRCCS-AOU San Martino-IST, Genova, Italy. aldo.pagano@unige.it.|Department of Experimental Medicine (DIMES), University of Genova, Largo Rosanna Benzi 10, 16132, Genova, Italy. aldo.pagano@unige.it.
authors.forename: Aldo
authors.initials: A
authors.lastname: Pagano
mesh_terms: D000200:Action Potentials / Q000502:physiology; D045744:Cell Line, Tumor; D006801:Humans; D009415:Nerve Net / Q000378:metabolism* / Q000473:pathology; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism*
chemical_list: C576282:NDM29 non-coding RNA, human; D022661:RNA, Untranslated
keywords: Cancer; Human neuronal network; Multi-electrode array (MEA); NDM29; Neuroblastoma; SKNBE2-S1.1