Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations.

Hdl Handle:
http://hdl.handle.net/10034/620627
Title:
Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations.
Authors:
Griffin, Sile M.; Pickard, Mark R.; Orme, Rowan P.; Hawkins, Clive P.; Williams, Adrian C.; Fricker, Rosemary A.
Abstract:
Vitamin B3 has been shown to play an important role during embryogenesis. Specifically, there is growing evidence that nicotinamide, the biologically active form of vitamin B3, plays a critical role as a morphogen in the differentiation of stem cells to mature cell phenotypes, including those of the central nervous system (CNS). Detailed knowledge of the action of small molecules during neuronal differentiation is not only critical for uncovering mechanisms underlying lineage-specification, but also to establish more effective differentiation protocols to obtain clinically relevant cells for regenerative therapies for neurodegenerative conditions such as Huntington's disease (HD). Thus, this study aimed to investigate the potential of nicotinamide to promote the conversion of stem cells to mature CNS neurons. METHODS: Nicotinamide was applied to differentiating mouse embryonic stem cells (mESC; Sox1GFP knock-in 46C cell line) during their conversion towards a neural fate. Cells were assessed for changes in their proliferation, differentiation and maturation; using immunocytochemistry and morphometric analysis methods. RESULTS: Results presented indicate that 10 mM nicotinamide, when added at the initial stages of differentiation, promoted accelerated progression of ESCs to a neural lineage in adherent monolayer cultures. By 14 days in vitro (DIV), early exposure to nicotinamide was shown to increase the numbers of differentiated βIII-tubulin-positive neurons. Nicotinamide decreased the proportion of pluripotent stem cells, concomitantly increasing numbers of neural progenitors at 4 DIV. These progenitors then underwent rapid conversion to neurons, observed by a reduction in Sox 1 expression and decreased numbers of neural progenitors in the cultures at 14 DIV. Furthermore, GABAergic neurons generated in the presence of nicotinamide showed increased maturity and complexity of neurites at 14 DIV. Therefore, addition of nicotinamide alone caused an accelerated passage of pluripotent cells through lineage specification and further to non-dividing mature neurons. CONCLUSIONS: Our results show that, within an optimal dose range, nicotinamide is able to singly and selectively direct the conversion of embryonic stem cells to mature neurons, and therefore may be a critical factor for normal brain development, thus supporting previous evidence of the fundamental role of vitamins and their metabolites during early CNS development. In addition, nicotinamide may offer a simple effective supplement to enhance the conversion of stem cells to clinically relevant neurons.
Affiliation:
Keele University; University of Chester; University Hospital of North Staffordshire; University of Birmingham
Citation:
Griffin, S. M., Pickard, M. R., Orme, R. P., Hawkins, C. P., Williams, A. C., & Fricker, R. A. (2017). Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations. PLoS One, 12(8), e0183358. https://doi.org/10.1371/journal.pone.0183358
Publisher:
Public Library of Science
Journal:
PLoS One
Publication Date:
17-Aug-2017
URI:
http://hdl.handle.net/10034/620627
DOI:
10.1371/journal.pone.0183358
Additional Links:
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0183358
Type:
Article
Language:
en
EISSN:
1932-6203
Appears in Collections:
Chester Medical School

Full metadata record

DC FieldValue Language
dc.contributor.authorGriffin, Sile M.en
dc.contributor.authorPickard, Mark R.en
dc.contributor.authorOrme, Rowan P.en
dc.contributor.authorHawkins, Clive P.en
dc.contributor.authorWilliams, Adrian C.en
dc.contributor.authorFricker, Rosemary A.en
dc.date.accessioned2017-09-25T13:17:39Z-
dc.date.available2017-09-25T13:17:39Z-
dc.date.issued2017-08-17-
dc.identifier.citationGriffin, S. M., Pickard, M. R., Orme, R. P., Hawkins, C. P., Williams, A. C., & Fricker, R. A. (2017). Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations. PLoS One, 12(8), e0183358. https://doi.org/10.1371/journal.pone.0183358en
dc.identifier.doi10.1371/journal.pone.0183358-
dc.identifier.urihttp://hdl.handle.net/10034/620627-
dc.description.abstractVitamin B3 has been shown to play an important role during embryogenesis. Specifically, there is growing evidence that nicotinamide, the biologically active form of vitamin B3, plays a critical role as a morphogen in the differentiation of stem cells to mature cell phenotypes, including those of the central nervous system (CNS). Detailed knowledge of the action of small molecules during neuronal differentiation is not only critical for uncovering mechanisms underlying lineage-specification, but also to establish more effective differentiation protocols to obtain clinically relevant cells for regenerative therapies for neurodegenerative conditions such as Huntington's disease (HD). Thus, this study aimed to investigate the potential of nicotinamide to promote the conversion of stem cells to mature CNS neurons. METHODS: Nicotinamide was applied to differentiating mouse embryonic stem cells (mESC; Sox1GFP knock-in 46C cell line) during their conversion towards a neural fate. Cells were assessed for changes in their proliferation, differentiation and maturation; using immunocytochemistry and morphometric analysis methods. RESULTS: Results presented indicate that 10 mM nicotinamide, when added at the initial stages of differentiation, promoted accelerated progression of ESCs to a neural lineage in adherent monolayer cultures. By 14 days in vitro (DIV), early exposure to nicotinamide was shown to increase the numbers of differentiated βIII-tubulin-positive neurons. Nicotinamide decreased the proportion of pluripotent stem cells, concomitantly increasing numbers of neural progenitors at 4 DIV. These progenitors then underwent rapid conversion to neurons, observed by a reduction in Sox 1 expression and decreased numbers of neural progenitors in the cultures at 14 DIV. Furthermore, GABAergic neurons generated in the presence of nicotinamide showed increased maturity and complexity of neurites at 14 DIV. Therefore, addition of nicotinamide alone caused an accelerated passage of pluripotent cells through lineage specification and further to non-dividing mature neurons. CONCLUSIONS: Our results show that, within an optimal dose range, nicotinamide is able to singly and selectively direct the conversion of embryonic stem cells to mature neurons, and therefore may be a critical factor for normal brain development, thus supporting previous evidence of the fundamental role of vitamins and their metabolites during early CNS development. In addition, nicotinamide may offer a simple effective supplement to enhance the conversion of stem cells to clinically relevant neurons.en
dc.language.isoenen
dc.publisherPublic Library of Scienceen
dc.relation.urlhttp://journals.plos.org/plosone/article?id=10.1371/journal.pone.0183358en
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectEmbryonic stem cellsen
dc.subjectNeuronsen
dc.subjectNicotinamideen
dc.subjectHuntington's diseaseen
dc.subjectBrain developmenten
dc.subjectVitaminsen
dc.titleNicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations.en
dc.typeArticleen
dc.identifier.eissn1932-6203-
dc.contributor.departmentKeele University; University of Chester; University Hospital of North Staffordshire; University of Birminghamen
dc.identifier.journalPLoS Oneen
dc.date.accepted2017-08-02-
or.grant.openaccessYesen
rioxxterms.funderInternally fundeden
rioxxterms.identifier.projectInternally funded research - Queen Elizabeth Hospital, Birmingham Charity funding and a Keele University ACORN studentshipen
rioxxterms.versionAMen
rioxxterms.licenseref.startdate2017-08-17-
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