title: In vitro differentiation of mouse embryonic stem cells into neurons of the dorsal forebrain.
abstract: Pluripotent embryonic stem cells (ESCs) are able to differentiate into all cell types in the organism including cortical neurons. To follow the dynamic generation of progenitors of the dorsal forebrain in vitro, we generated ESCs from D6-GFP mice in which GFP marks neocortical progenitors and neurons after embryonic day (E) 10.5. We used several cell culture protocols for differentiation of ESCs into progenitors and neurons of the dorsal forebrain. In cell culture, GFP-positive cells were induced under differentiation conditions in quickly formed embryoid bodies (qEBs) after 10-12 day incubation. Activation of Wnt signaling during ESC differentiation further stimulated generation of D6-GFP-positive cortical cells. In contrast, differentiation protocols using normal embryoid bodies (nEBs) yielded only a few D6-GFP-positive cells. Gene expression analysis revealed that multiple components of the canonical Wnt signaling pathway were expressed during the development of embryoid bodies. As shown by immunohistochemistry and quantitative qRT-PCR, D6-GFP-positive cells from qEBs expressed genes that are characteristic for the dorsal forebrain such as Pax6, Dach1, Tbr1, Tbr2, or Sox5. qEBs culture allowed the formation of a D6-GFP positive pseudo-polarized neuroepithelium with the characteristic presence of N-cadherin at the apical pole resembling the structure of the developing neocortex.
authors.affiliation: School of Medicine, Zhengzhou University, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Jing
authors.forename: Ondrej
authors.initials: O
authors.lastname: Machon
authors.forename: Ales
authors.initials: A
authors.lastname: Hampl
authors.forename: Petr
authors.initials: P
authors.lastname: Dvorak
authors.forename: Ying
authors.initials: Y
authors.lastname: Xing
authors.forename: Stefan
authors.initials: S
authors.lastname: Krauss
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002449:Cell Aggregation; D002454:Cell Differentiation; D016616:Cryoultramicrotomy; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009928:Organ Specificity; D038081:Organogenesis; D016548:Prosencephalon / Q000166:cytology* / Q000378:metabolism; D015398:Signal Transduction; D013687:Telencephalon / Q000166:cytology / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism
chemical_list: D015415:Biomarkers; D051153:Wnt Proteins; D049452:Green Fluorescent Proteins