Hdl Handle:
http://hdl.handle.net/10675.2/61
Title:
Directed neuronal differentiation of human embryonic stem cells.
Authors:
Schulz, Thomas C; Palmarini, Gail M; Noggle, Scott A; Weiler, Deborah A; Mitalipova, Maisam M; Condie, Brian G.
Abstract:
BACKGROUND: We have developed a culture system for the efficient and directed differentiation of human embryonic stem cells (HESCs) to neural precursors and neurons.HESC were maintained by manual passaging and were differentiated to a morphologically distinct OCT-4+/SSEA-4- monolayer cell type prior to the derivation of embryoid bodies. Embryoid bodies were grown in suspension in serum free conditions, in the presence of 50% conditioned medium from the human hepatocarcinoma cell line HepG2 (MedII). RESULTS: A neural precursor population was observed within HESC derived serum free embryoid bodies cultured in MedII conditioned medium, around 7-10 days after derivation. The neural precursors were organized into rosettes comprised of a central cavity surrounded by ring of cells, 4 to 8 cells in width. The central cells within rosettes were proliferating, as indicated by the presence of condensed mitotic chromosomes and by phosphoHistone H3 immunostaining. When plated and maintained in adherent culture, the rosettes of neural precursors were surrounded by large interwoven networks of neurites. Immunostaining demonstrated the expression of nestin in rosettes and associated non-neuronal cell types, and a radial expression of Map-2 in rosettes. Differentiated neurons expressed the markers Map-2 and Neurofilament H, and a subpopulation of the neurons expressed tyrosine hydroxylase, a marker for dopaminergic neurons. CONCLUSION: This novel directed differentiation approach led to the efficient derivation of neuronal cultures from HESCs, including the differentiation of tyrosine hydroxylase expressing neurons. HESC were morphologically differentiated to a monolayer OCT-4+ cell type, which was used to derive embryoid bodies directly into serum free conditions. Exposure to the MedII conditioned medium enhanced the derivation of neural precursors, the first example of the effect of this conditioned medium on HESC.
Citation:
BMC Neurosci. 2003 Oct 22; 4:27
Issue Date:
10-May-2004
URI:
http://hdl.handle.net/10675.2/61
DOI:
10.1186/1471-2202-4-27
PubMed ID:
14572319
PubMed Central ID:
PMC272931
Type:
Journal Article; Research Support, Non-U.S. Gov't
ISSN:
1471-2202
Appears in Collections:
Institute of Molecular Medicine and Genetics: Faculty Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorSchulz, Thomas Cen_US
dc.contributor.authorPalmarini, Gail Men_US
dc.contributor.authorNoggle, Scott Aen_US
dc.contributor.authorWeiler, Deborah Aen_US
dc.contributor.authorMitalipova, Maisam Men_US
dc.contributor.authorCondie, Brian G.en_US
dc.date.accessioned2010-09-24T21:26:50Z-
dc.date.available2010-09-24T21:26:50Z-
dc.date.issued2004-05-10en_US
dc.identifier.citationBMC Neurosci. 2003 Oct 22; 4:27en_US
dc.identifier.issn1471-2202en_US
dc.identifier.pmid14572319en_US
dc.identifier.doi10.1186/1471-2202-4-27en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/61-
dc.description.abstractBACKGROUND: We have developed a culture system for the efficient and directed differentiation of human embryonic stem cells (HESCs) to neural precursors and neurons.HESC were maintained by manual passaging and were differentiated to a morphologically distinct OCT-4+/SSEA-4- monolayer cell type prior to the derivation of embryoid bodies. Embryoid bodies were grown in suspension in serum free conditions, in the presence of 50% conditioned medium from the human hepatocarcinoma cell line HepG2 (MedII). RESULTS: A neural precursor population was observed within HESC derived serum free embryoid bodies cultured in MedII conditioned medium, around 7-10 days after derivation. The neural precursors were organized into rosettes comprised of a central cavity surrounded by ring of cells, 4 to 8 cells in width. The central cells within rosettes were proliferating, as indicated by the presence of condensed mitotic chromosomes and by phosphoHistone H3 immunostaining. When plated and maintained in adherent culture, the rosettes of neural precursors were surrounded by large interwoven networks of neurites. Immunostaining demonstrated the expression of nestin in rosettes and associated non-neuronal cell types, and a radial expression of Map-2 in rosettes. Differentiated neurons expressed the markers Map-2 and Neurofilament H, and a subpopulation of the neurons expressed tyrosine hydroxylase, a marker for dopaminergic neurons. CONCLUSION: This novel directed differentiation approach led to the efficient derivation of neuronal cultures from HESCs, including the differentiation of tyrosine hydroxylase expressing neurons. HESC were morphologically differentiated to a monolayer OCT-4+ cell type, which was used to derive embryoid bodies directly into serum free conditions. Exposure to the MedII conditioned medium enhanced the derivation of neural precursors, the first example of the effect of this conditioned medium on HESC.en_US
dc.rightsThe PMC Open Access Subset is a relatively small part of the total collection of articles in PMC. Articles in the PMC Open Access Subset are still protected by copyright, but are made available under a Creative Commons or similar license that generally allows more liberal redistribution and reuse than a traditional copyrighted work. Please refer to the license statement in each article for specific terms of use. The license terms are not identical for all articles in this subset.en_US
dc.subject.meshAnimalsen_US
dc.subject.meshAntigens, Differentiation / biosynthesisen_US
dc.subject.meshCell Culture Techniques / methodsen_US
dc.subject.meshCell Differentiation / drug effects / physiologyen_US
dc.subject.meshCell Division / drug effects / physiologyen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshCoculture Techniques / methodsen_US
dc.subject.meshCulture Media, Conditioned / pharmacologyen_US
dc.subject.meshCulture Media, Serum-Free / pharmacologyen_US
dc.subject.meshFibroblasts / cytology / metabolismen_US
dc.subject.meshHumansen_US
dc.subject.meshMiceen_US
dc.subject.meshNeurons / cytology / drug effects / metabolismen_US
dc.subject.meshStem Cells / cytology / drug effects / physiologyen_US
dc.subject.meshTime Factorsen_US
dc.titleDirected neuronal differentiation of human embryonic stem cells.en_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
dc.identifier.pmcidPMC272931en_US
dc.contributor.corporatenameInstitute of Molecular Medicine and Geneticsen_US

Related articles on PubMed

All Items in Scholarly Commons are protected by copyright, with all rights reserved, unless otherwise indicated.