title: Three dimensional cellular microarray platform for human neural stem cell differentiation and toxicology.
abstract: We developed a three-dimensional (3D) cellular microarray platform for the high-throughput (HT) analysis of human neural stem cell (hNSC) growth and differentiation. The growth of an immortalized hNSC line, ReNcell VM, was evaluated on a miniaturized cell culture chip consisting of 60nl spots of cells encapsulated in alginate, and compared to standard 2D well plate culture conditions. Using a live/dead cell viability assay, we demonstrated that the hNSCs are able to expand on-chip, albeit with lower proliferation rates and viabilities than in conventional 2D culture platforms. Using an in-cell, on-chip immunofluorescence assay, which provides quantitative information on cellular levels of proteins involved in neural fate, we demonstrated that ReNcell VM can preserve its multipotent state during on-chip expansion. Moreover, differentiation of the hNSCs into glial progeny was achieved both off- and on-chip six days after growth factor removal, accompanied by a decrease in the neural progenitor markers. The versatility of the platform was further demonstrated by complementing the cell culture chip with a chamber system that allowed us to screen for differential toxicity of small molecules to hNSCs. Using this approach, we showed differential toxicity when evaluating three neurotoxic compounds and one antiproliferative compound, and the null effect of a non-toxic compound at relevant concentrations. Thus, our 3D high-throughput microarray platform may help predict, in vitro, which compounds pose an increased threat to neural development and should therefore be prioritized for further screening and evaluation.
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, USA.
authors.forename: Luciana
authors.initials: L
authors.lastname: Meli
authors.affiliation: Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, University of Lisbon, Portugal; Department of Bioengineering, Instituto Superior Técnico, University of Lisbon, Portugal.
authors.forename: Hélder S C
authors.initials: HS
authors.lastname: Barbosa
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, USA.
authors.forename: Anne Marie
authors.initials: AM
authors.lastname: Hickey
authors.affiliation: Department of Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA.
authors.forename: Leyla
authors.initials: L
authors.lastname: Gasimli
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Nierode
authors.affiliation: Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, University of Lisbon, Portugal; Department of Bioengineering, Instituto Superior Técnico, University of Lisbon, Portugal.
authors.forename: Maria Margarida
authors.initials: MM
authors.lastname: Diogo
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, USA; Department of Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA; Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA; Department of Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
authors.affiliation: Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, University of Lisbon, Portugal; Department of Bioengineering, Instituto Superior Técnico, University of Lisbon, Portugal.
authors.forename: Joaquim M S
authors.initials: JM
authors.lastname: Cabral
authors.affiliation: Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY, USA; Department of Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA; Department of Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA; Department of Materials Science and Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA. Electronic address: dordick@rpi.edu.
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Dordick
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D058953:Neural Stem Cells / Q000166:cytology*; D009457:Neuroglia / Q000166:cytology