title: Neural differentiation of pluripotent cells in 3D alginate-based cultures.
abstract: Biomaterial-supported culture methods, allowing for directed three-dimensional differentiation of stem cells are an alternative to canonical two-dimensional cell cultures. In this paper, we evaluate the suitability of alginate for three-dimensional cultures to enhance differentiation of mouse embryonic stem cells (mESCs) towards neural lineages. We tested whether encapsulation of mESCs within alginate beads could support and/or enhance neural differentiation with respect to two-dimensional cultures. We encapsulated cells in beads of alginate with or without modification by fibronectin (Fn) or hyaluronic acid (HA). Gene expression analysis showed that cells grown in alginate and alginate-HA present increased differentiation toward neural lineages with respect to the two-dimensional control and to Fn group. Immunocytochemistry analyses confirmed these results, further showing terminal differentiation of neurons as seen by the expression of synaptic markers and markers of different neuronal subtypes. Our data show that alginate, alone or modified, is a suitable biomaterial to promote in vitro differentiation of pluripotent cells toward neural fates.
authors.affiliation: Centre for Integrative Biology (CIBIO), University of Trento, 38123 Trento, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Bozza
authors.affiliation: Fischell Department of Bioengineering, University of Maryland, 20742 Maryland, USA.
authors.forename: Emily E
authors.initials: EE
authors.lastname: Coates
authors.affiliation: Centre for Integrative Biology (CIBIO), University of Trento, 38123 Trento, Italy.
authors.forename: Tania
authors.initials: T
authors.lastname: Incitti
authors.affiliation: Fischell Department of Bioengineering, University of Maryland, 20742 Maryland, USA.
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Ferlin
authors.affiliation: Centre for Integrative Biology (CIBIO), University of Trento, 38123 Trento, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Messina
authors.affiliation: CNR Neuroscience Institute, National Research Council (CNR), 20133 Milan, Italy; Humanitas Clinical and Research Center, 20133 Milan, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Menna
authors.affiliation: Centre for Integrative Biology (CIBIO), University of Trento, 38123 Trento, Italy; CNR Neuroscience Institute, National Research Council (CNR), 56124 Pisa, Italy.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Bozzi
authors.affiliation: Fischell Department of Bioengineering, University of Maryland, 20742 Maryland, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Fisher
authors.affiliation: Centre for Integrative Biology (CIBIO), University of Trento, 38123 Trento, Italy; CNR Neuroscience Institute, National Research Council (CNR), 56124 Pisa, Italy. Electronic address: casarosa@science.unitn.it.
authors.forename: Simona
authors.initials: S
authors.lastname: Casarosa
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006820:Hyaluronic Acid / Q000737:chemistry; D051379:Mice; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000464:Alginates; D001672:Biocompatible Materials; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D006820:Hyaluronic Acid
keywords: Alginate; Hyaluronic acid; Neural differentiation; Pluripotent cells; Three dimensional cultures