In Vitro Differentiation of Mouse Embryonic Stem Cells into Neurons of the Dorsal Forebrain

JING, Ying, Ondřej MACHOŇ, Aleš HAMPL, Petr DVOŘÁK, Ying XING and Stefan KRAUSS. In Vitro Differentiation of Mouse Embryonic Stem Cells into Neurons of the Dorsal Forebrain. Cellular and Molecular Neurobiology. Springer, 2011, vol. 31, No 5, p. 715-727. ISSN 0272-4340. Available from: https://dx.doi.org/10.1007/s10571-011-9669-2.

Basic information

• Original name: In Vitro Differentiation of Mouse Embryonic Stem Cells into Neurons of the Dorsal Forebrain
• Authors: JING, Ying (156 China), Ondřej MACHOŇ (203 Czech Republic), Aleš HAMPL (203 Czech Republic, belonging to the institution), Petr DVOŘÁK (203 Czech Republic, belonging to the institution), Ying XING (156 China) and Stefan KRAUSS (578 Norway, guarantor).
• Edition: Cellular and Molecular Neurobiology, Springer, 2011, 0272-4340.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 30000 3. Medical and Health Sciences
• Country of publisher: United States of America
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 1.969
• RIV identification code: RIV/00216224:14110/11:00052566
• Organization unit: Faculty of Medicine
• Doi: http://dx.doi.org/10.1007/s10571-011-9669-2
• UT WoS: 000290914300008
• Keywords in English: Mouse; embryonic stem cell; Embryonic forebrain; Stem cell; differentiation; Wnt signaling
• Tags: International impact

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-GFPpositive 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.