title: Efficient neuronal in vitro and in vivo differentiation after immunomagnetic purification of mESC derived neuronal precursors.
abstract: The cellular heterogeneity that is generated during the differentiation of pluripotent stem cells into specific neural subpopulations represents a major obstacle for experimental and clinical progress. To address this problem we developed an optimized strategy for magnetic isolation of PSA-NCAM positive neuronal precursors from embryonic stem cells (ESCs) derived neuronal cultures. PSA-NCAM enrichment at an early step of the in vitro differentiation process increased the number of ES cell derived neurons and reduced cellular diversity. Gene expression analysis revealed that mainly genes involved in neuronal activity were over-represented after purification. In vitro derived PSA-NCAM(+) enriched precursors were characterized in vivo through grafting into the forebrain of adult mice. While unsorted control cells 40 days post graft gave rise to a mixed population composed of immature precursors, early postmitotic neurons and glial cells, PSA-NCAM(+) enriched cells differentiated predominantly into NeuN positive cells. Furthermore, PSA-NCAM enriched population showed efficient migration towards the olfactory bulb after transplantation into the rostral migratory stream of the forebrain neurogenic system. Thus, enrichment of neuronal precursors based on PSA-NCAM expression represents a general and straightforward approach to narrow cellular heterogeneity during neuronal differentiation of pluripotent cells.
authors.affiliation: Miltenyi Biotec GmbH, 51429 Bergisch Gladbach, Germany. serena.barral@univ-amu.fr
authors.forename: Serena
authors.initials: S
authors.lastname: Barral
authors.forename: Josephine
authors.initials: J
authors.lastname: Ecklebe
authors.forename: Stefan
authors.initials: S
authors.lastname: Tomiuk
authors.forename: Marie-Catherine
authors.initials: MC
authors.lastname: Tiveron
authors.forename: Angélique
authors.initials: A
authors.lastname: Desoeuvre
authors.forename: Dominik
authors.initials: D
authors.lastname: Eckardt
authors.forename: Harold
authors.initials: H
authors.lastname: Cremer
authors.forename: Andreas
authors.initials: A
authors.lastname: Bosio
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018189:Immunomagnetic Separation; D051379:Mice; D008810:Mice, Inbred C57BL; D039842:Neural Cell Adhesion Molecule L1 / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D012794:Sialic Acids / Q000378:metabolism; D033581:Stem Cell Transplantation
chemical_list: D000199:Actins; D039842:Neural Cell Adhesion Molecule L1; D012794:Sialic Acids; C485184:enhanced green fluorescent protein; C103469:polysialyl neural cell adhesion molecule; D049452:Green Fluorescent Proteins