title: Glycoconjugates reveal diversity of human neural stem cells (hNSCs) derived from human induced pluripotent stem cells (hiPSCs).
abstract: Neural stem cells (NSCs) have the ability to self-renew and to differentiate into various cell types of the central nervous system. This potential can be recapitulated by human induced pluripotent stem cells (hiPSCs) in vitro. The differentiation capacity of hiPSCs is characterized by several stages with distinct morphologies and the expression of various marker molecules. We used the monoclonal antibodies (mAbs) 487LeX, 5750LeXand 473HD to analyze the expression pattern of particular carbohydrate motifs as potential markers at six differentiation stages of hiPSCs. Mouse ESCs were used as a comparison. At the pluripotent stage, 487LeX-, 5750LeX- and 473HD-related glycans were differently expressed. Later, cells of the three germ layers in embryoid bodies (hEBs) and, even after neuralization of hEBs, subpopulations of cells were labeled with these surface antibodies. At the human rosette-stage of NSCs (hR-NSC), LeX- and 473HD-related epitopes showed antibody-specific expression patterns. We also found evidence that these surface antibodies could be used to distinguish the hR-NSCs from the hSR-NSCs stages. Characterization of hNSCsFGF-2/EGFderived from hSR-NSCs revealed that both LeX antibodies and the 473HD antibody labeled subpopulations of hNSCsFGF-2/EGF. Finally, we identified potential LeX carrier molecules that were spatiotemporally regulated in early and late stages of differentiation. Our study provides new insights into the regulation of glycoconjugates during early human stem cell development. The mAbs 487LeX, 5750LeXand 473HD are promising tools for identifying distinct stages during neural differentiation.
authors.affiliation: Department of Cell Morphology and Molecular Neurobiology, Ruhr University Bochum, Bochum, Germany.
authors.forename: Majury
authors.initials: M
authors.lastname: Kandasamy
authors.affiliation: Department of Cell Morphology and Molecular Neurobiology, Ruhr University Bochum, Bochum, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Roll
authors.affiliation: Department of Cell Morphology and Molecular Neurobiology, Ruhr University Bochum, Bochum, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Langenstroth
authors.affiliation: Institute of Reconstructive Neurobiology, LIFE & BRAIN Center, University of Bonn, Bonn, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
authors.affiliation: Department of Cell Morphology and Molecular Neurobiology, Ruhr University Bochum, Bochum, Germany. andreas.faissner@rub.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Faissner
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000943:Antigens, Differentiation / Q000096:biosynthesis; D002454:Cell Differentiation; D002460:Cell Line; D016764:Cell Polarity; D000939:Epitopes / Q000096:biosynthesis; D006001:Glycoconjugates / Q000096:biosynthesis / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000276:immunology; D011134:Polysaccharides / Q000096:biosynthesis
chemical_list: D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000939:Epitopes; D006001:Glycoconjugates; D011134:Polysaccharides
keywords: 473HD epitope; Cell surface glycans; Chondroitin sulfate proteoglycan; LewisX-type glycans; Phosphacan