title: High-purity lineage selection of embryonic stem cell-derived neurons.
abstract: The derivation of somatic cell types from pluripotent and self-renewing embryonic stem (ES) cells offers attractive prospects for basic research, compound development, and regenerative medicine. A key prerequisite for biomedical applications of ES cells is the ability to differentiate and isolate defined somatic cell populations at high purity. In this study, we explore the potential of the Talpha1- enhanced green fluorescent protein (EGFP) transgene and polysialic acid (PSA)-neural cell adhesion molecule (NCAM) as lineage selection markers for the derivation of ES cell-derived neurons. Upon controlled in vitro differentiation, ES cells engineered to express EGFP under control of the Talpha1-tubulin promoter exhibited exclusive transgene expression in neurons. Similarly, PSA-NCAM expression during the early stages of ES cell differentiation was restricted to neuronal progeny. Talpha1- EGFP- and PSA-NCAM-positive neurons comprised both inhibitory and excitatory phenotypes. Compared to Talpha1-EGFP, the expression of PSA-NCAM was initiated at slightly earlier stages of neural differentiation. FACSorting of Talpha1-EGFP-positive cells and immunopanning of PSA-NCAMexpressing cells yielded neuronal populations at purities up to 99.6% and 96.9%, respectively. These findings depict Talpha1-EGFP and PSA-NCAM as suitable markers for high-purity selection of early ES cell-derived neurons.
authors.affiliation: Institute of Reconstructive Neurobiology, University of Bonn Medical Center and Hertie Foundation, Bonn, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Schmandt
authors.forename: Eybe
authors.initials: E
authors.lastname: Meents
authors.forename: Gudrun
authors.initials: G
authors.lastname: Gossrau
authors.forename: Volker
authors.initials: V
authors.lastname: Gornik
authors.forename: Shigeo
authors.initials: S
authors.lastname: Okabe
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods* / Q000592:standards; D004622:Embryo, Mammalian / Q000166:cytology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D019006:Neural Cell Adhesion Molecules / Q000032:analysis; D009474:Neurons / Q000166:cytology*; D010641:Phenotype; D012794:Sialic Acids / Q000032:analysis; D013234:Stem Cells / Q000166:cytology*; D014404:Tubulin / Q000235:genetics
chemical_list: D019006:Neural Cell Adhesion Molecules; D012794:Sialic Acids; D014404:Tubulin; C021319:polysialic acid; D049452:Green Fluorescent Proteins