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Can manipulation of differentiation conditions eliminate proliferative cells from a population of ES cell-derived forebrain cells?

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Author
Precious, Sophie V.
Kelly, Claire
Allen, Nicholas D.
Rosser, Anne E.
Date
2016-01-11
Type
Article
Publisher
Taylor Francis
Metadata
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Abstract
There is preliminary evidence that implantation of primary fetal striatal cells provides functional benefit in patients with Huntington’s disease, a neurodegenerative condition resulting in loss of medium-sized spiny neurons (MSN) of the striatum. Scarcity of primary fetal tissue means it is important to identify a renewable source of cells from which to derive donor MSNs. Embryonic stem (ES) cells, which predominantly default to telencephalic-like precursors in chemically defined medium (CDM), offer a potentially inexhaustible supply of cells capable of generating the desired neurons. Using an ES cell line, with the forebrain marker FoxG1 tagged to the LacZ reporter, we assessed effects of known developmental factors on the yield of forebrain-like precursor cells in CDM suspension culture. Addition of FGF2, but not DKK1, increased the proportion of FoxG1-expressing cells at day 8 of neural induction. Oct4 was expressed at day 8, but was undetectable by day 16. Differentiation of day 16 precursors generated GABA-expressing neurons, with few DARPP32 positive MSNs. Transplantation of day 8 precursor cells into quinolinic acid-lesioned striata resulted in generation of teratomas. However, transplantation of day 16 precursors yielded grafts expressing neuronal markers including NeuN, calbindin and parvalbumin, but no DARPP32 6 weeks post-transplantation. Manipulation of fate of ES cells requires optimization of both concentration and timing of addition of factors to culture systems to generate the desired phenotypes. Furthermore, we highlight the value of increasing the precursor phase of ES cell suspension culture when directing differentiation towards forebrain fate, so as to dramatically reduce the risk of teratoma formation.
Journal/conference proceeding
Neurogenesis
Citation
Precious S.V., Kelly, C.M., Allen, N.D. and Rosser, A.E. (2016) 'Can manipulation of differentiation conditions eliminate proliferative cells from a population of ES cell-derived forebrain cells?', Neurogenesis
URI
http://hdl.handle.net/10369/7730
DOI
http://dx.doi.org/10.1080/23262133.2015.1127311
Description
This article was published in Neurogenesis on 11th January 2016 (online) available at http://dx.doi.org/10.1080/23262133.2015.1127311
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