title: Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation.
abstract: Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (iPSCs) are traditionally maintained and proliferated utilizing two-dimensional (2D) adherent monolayer culture systems. However, NPCs cultured using this system hardly reflect the intrinsic spatial development of brain tissue. In this study, we determined that culturing iPSC-derived NPCs as three-dimensional (3D) floating neurospheres resulted in increased expression of the neural progenitor cell (NPC) markers, PAX6 and NESTIN. Expansion of NPCs in 3D culture methods also resulted in a more homogenous PAX6 expression when compared to 2D culture methods. Furthermore, the 3D propagation method for NPCs resulted in a significant higher expression of the astrocyte markers  GFAP and aquaporin 4 (AQP4) in the differentiated cells. Thus, our 3D propagation method could constitute a useful tool to promote NPC homogeneity and also to increase the differentiation potential of iPSC towards astrocytes.
authors.affiliation: Department of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, Denmark.
authors.forename: Shuling
authors.initials: S
authors.lastname: Zhou
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary.
authors.forename: Karolina
authors.initials: K
authors.lastname: Szczesna
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary; Molecular Animal Biotechnology Laboratory, SZIE, Gödöllo 2100, Hungary.
authors.forename: Anna
authors.initials: A
authors.lastname: Ochalek
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary.
authors.forename: Julianna
authors.initials: J
authors.lastname: Kobolák
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary.
authors.forename: Eszter
authors.initials: E
authors.lastname: Varga
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary.
authors.forename: Csilla
authors.initials: C
authors.lastname: Nemes
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary.
authors.forename: Abinaya
authors.initials: A
authors.lastname: Chandrasekaran
authors.affiliation: Bioneer A/S, 2970 Hørsholm, Denmark.
authors.forename: Mikkel
authors.initials: M
authors.lastname: Rasmussen
authors.affiliation: Department of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, Denmark.
authors.forename: Susanna
authors.initials: S
authors.lastname: Cirera
authors.affiliation: Department of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, Denmark.
authors.forename: Poul
authors.initials: P
authors.lastname: Hyttel
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary; Molecular Animal Biotechnology Laboratory, SZIE, Gödöllo 2100, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Dinnyés
authors.affiliation: Department of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, Denmark.
authors.forename: Kristine K
authors.initials: KK
authors.lastname: Freude
authors.affiliation: BioTalentum Ltd., Gödöllo 2100, Hungary.
authors.forename: Hasan X
authors.initials: HX
authors.lastname: Avci