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Robust induction of DARPP32-expressing GABAergic striatal neurons from human pluripotent stem cells

Fjodorova, Marija and Li, Meng ORCID: 2018. Robust induction of DARPP32-expressing GABAergic striatal neurons from human pluripotent stem cells. Precious, Sophie, Rosser, Anne and Dunnett, Stephen, eds. Huntington’s Disease, Vol. 1780. Methods in Molecular Biology, New York: Humana Press, pp. 585-605. (10.1007/978-1-4939-7825-0_27)

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Efficient generation of disease relevant neuronal subtypes from human pluripotent stem cells (PSCs) is fundamental for realizing their promise in disease modeling, pharmaceutical drug screening and cell therapy. Here we describe a step-by-step protocol for directing the differentiation of human embryonic and induced PSCs (hESCs and hiPSCs, respectively) toward medium spiny neurons, the type of cells that are preferentially lost in Huntington’s disease patients. This method is based on a novel concept of Activin A-dependent induction of the lateral ganglionic/striatal fate using a simple monolayer culture paradigm under chemically defined conditions. Transplantable medium spiny neuron progenitors amenable for cryopreservation are produced in less than 20 days, which differentiate and mature into a high yield of dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDa (DARPP32) expressing gamma-aminobutyric acid (GABA)-ergic neurons in vitro and in the adult rat brain after transplantation. This method has been validated in multiple hESC and hiPSC lines, and is independent of the regime for PSC maintenance.

Item Type: Book Section
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: Humana Press
ISBN: 9781493978243
ISSN: 1064-3745
Date of First Compliant Deposit: 4 January 2019
Date of Acceptance: 1 March 2017
Last Modified: 06 Nov 2023 19:31

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