title: Screening of bioactive peptides using an embryonic stem cell-based neurodifferentiation assay.
abstract: Differentiation of pluripotent stem cells, PSCs, towards neural lineages has attracted significant attention, given the potential use of such cells for in vitro studies and for regenerative medicine. The present experiments were designed to identify bioactive peptides which direct PSC differentiation towards neural cells. Fifteen peptides were designed based on NCAM, FGFR, and growth factors sequences. The effect of peptides was screened using a mouse embryonic stem cell line expressing luciferase dual reporter construct driven by promoters for neural tubulin and for elongation factor 1. Cell number was estimated by measuring total cellular DNA. We identified five peptides which enhanced activities of both promoters without relevant changes in cell number. We selected the two most potent peptides for further analysis: the NCAM-derived mimetic FGLL and the synthetic NCAM ligand, Plannexin. Both compounds induced phenotypic neuronal differentiation, as evidenced by increased neurite outgrowth. In summary, we used a simple, but sensitive screening approach to identify the neurogenic peptides. These peptides will not only provide new clues concerning pathways of neurogenesis, but they may also be interesting biotechnology tools for in vitro generation of neurons.
authors.affiliation: ENKAM Pharmaceuticals A/S, Copenhagen, Denmark.
authors.forename: Ruodan
authors.initials: R
authors.lastname: Xu
authors.forename: Maxime
authors.initials: M
authors.lastname: Feyeux
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Julien
authors.forename: Csilla
authors.initials: C
authors.lastname: Nemes
authors.forename: Morten
authors.initials: M
authors.lastname: Albrechtsen
authors.forename: Andras
authors.initials: A
authors.lastname: Dinnyés
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Krause
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D051379:Mice; D058953:Neural Stem Cells / Q000187:drug effects*; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure; D010455:Peptides / Q000494:pharmacology*
chemical_list: D007202:Indicators and Reagents; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides