title: Functional and phenotypic differences of pure populations of stem cell-derived astrocytes and neuronal precursor cells.
abstract: Availability of homogeneous astrocyte populations would facilitate research concerning cell plasticity (metabolic and transcriptional adaptations; innate immune responses) and cell cycle reactivation. Current protocols to prepare astrocyte cultures differ in their final content of immature precursor cells, preactivated cells or entirely different cell types. A new method taking care of all these issues would improve research on astrocyte functions. We found here that the exposure of a defined population of pluripotent stem cell-derived neural stem cells (NSC) to BMP4 results in pure, nonproliferating astrocyte cultures within 24-48 h. These murine astrocytes generated from embryonic stem cells (mAGES) expressed the positive markers GFAP, aquaporin 4 and GLT-1, supported neuronal function, and acquired innate immune functions such as the response to tumor necrosis factor and interleukin 1. The protocol was applicable to several normal or disease-prone pluripotent cell lines, and the corresponding mAGES all exited the cell cycle and lost most of their nestin expression, in contrast to astrocytes generated by serum-addition or obtained as primary cultures. Comparative gene expression analysis of mAGES and NSC allowed quantification of differences between the two cell types and a definition of an improved marker set to define astrocytes. Inclusion of several published data sets in this transcriptome comparison revealed the similarity of mAGES with cortical astrocytes in vivo. Metabolic analysis of homogeneous NSC and astrocyte populations revealed distinct neurochemical features: both cell types synthesized glutamine and citrate, but only mature astrocytes released these metabolites. Thus, the homogeneous cultures allowed an improved definition of NSC and astrocyte features.
authors.affiliation: The Doerenkamp-Zbinden Chair of in-Vitro Toxicology and Biomedicine/Alternatives to Animal Experimentation, University of Konstanz, Konstanz, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Kleiderman
authors.affiliation: IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: João V
authors.initials: JV
authors.lastname: Sá
authors.affiliation: IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Ana P
authors.initials: AP
authors.lastname: Teixeira
authors.affiliation: IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Brito
authors.affiliation: The Doerenkamp-Zbinden Chair of in-Vitro Toxicology and Biomedicine/Alternatives to Animal Experimentation, University of Konstanz, Konstanz, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Gutbier
authors.affiliation: Department of Earth Science, University of Bergen, Allégaten 41, 5007 Bergen, Norway.
authors.forename: Lars G
authors.initials: LG
authors.lastname: Evje
authors.affiliation: Department of Pharmacy, College of Health Sciences, Mekelle University, Tigray, Ethiopia.
authors.forename: Mussie G
authors.initials: MG
authors.lastname: Hadera
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Belvaux, L-4366, Luxembourg.
authors.forename: Enrico
authors.initials: E
authors.lastname: Glaab
authors.affiliation: Institute of Neurophysiology and Center for Molecular Medicine, Cologne (CMMC), University of Cologne, Cologne, Germany.
authors.forename: Margit
authors.initials: M
authors.lastname: Henry
authors.affiliation: Institute of Neurophysiology and Center for Molecular Medicine, Cologne (CMMC), University of Cologne, Cologne, Germany.
authors.forename: Agapios
authors.initials: A
authors.lastname: Sachinidis
authors.affiliation: IBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal.|Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, Copenhagen, Denmark.|Department of Neuroscience, Norwegian University of Science and Technology, Faculty of Medicine, Trondheim, Norway.
authors.forename: Ursula
authors.initials: U
authors.lastname: Sonnewald
authors.affiliation: The Doerenkamp-Zbinden Chair of in-Vitro Toxicology and Biomedicine/Alternatives to Animal Experimentation, University of Konstanz, Konstanz, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Leist
mesh_terms: D000818:Animals; D051401:Aquaporin 4 / Q000235:genetics / Q000378:metabolism; D001253:Astrocytes / Q000502:physiology*; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000502:physiology*; D027342:Excitatory Amino Acid Transporter 2 / Q000235:genetics / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D064231:Nestin / Q000378:metabolism; D058953:Neural Stem Cells / Q000502:physiology*; D013997:Time Factors; D059467:Transcriptome / Q000502:physiology
chemical_list: D051401:Aquaporin 4; C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D016207:Cytokines; D027342:Excitatory Amino Acid Transporter 2; D005904:Glial Fibrillary Acidic Protein; D019394:Ki-67 Antigen; D064231:Nestin; C528900:glial fibrillary astrocytic protein, mouse; D019344:Lactic Acid; D018698:Glutamic Acid; D005947:Glucose
keywords: astrocytes; differentiation; metabolic flux; neural stem cells; transcriptome